Compounds useful for treating disorders related to ret

ABSTRACT

Described herein are compounds that inhibit wild-type RET and its resistant mutants, pharmaceutical compositions including such compounds, and methods of using such compounds and compositions.

CLAIM OF PRIORITY

This application claims priority from U.S. Provisional Application No.62/365,724, filed Jul. 22, 2016, which is incorporated herein in itsentirety.

This disclosure relates to inhibitors of RET that are active againstwild-type RET and its resistant mutants.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been filedelectronically in ASCII format and is hereby incorporated by referencein its entirety. Said ASCII copy, created on Jul. 21, 2017, is named14320_0019-00000_SL.txt and is 32,335 bytes in size.

BACKGROUND

RET is a receptor tyrosine kinase that activates multiple downstreampathways involved in cell proliferation and survival. RET fusions areimplicated in several cancers including papillary thyroid carcinoma andnon-small cell lung cancer. A genomics analysis on the landscape ofkinase fusions identified RET fusions in breast and colon cancer patientsamples, providing therapeutic rationale for the use of RET inhibitorsin multiple patient subpopulations.

The identification of RET fusions as drivers in some cancers promptedthe use of approved multi-kinase inhibitors with RET inhibitory activityto treat patients whose tumors express a RET fusion protein. However,these drugs cannot always be dosed at the levels required tosufficiently inhibit RET due to toxicities that result from inhibitionof targets other than RET. Further, one of the greatest challenges intreating cancer is the ability of tumor cells to become resistant totherapy. Kinase reactivation via mutation is a common mechanism ofresistance. When resistance occurs, the patient's treatment options areoften very limited, and the cancer progresses, unchecked, in mostinstances. There is thus a need for compounds that inhibit RET, as wellas its resistant mutants.

SUMMARY

In one aspect, the disclosure features a compound of Formula (I) or apharmaceutically acceptable salt thereof, wherein:

or a pharmaceutically acceptable salt thereof, wherein: ring A is anaryl or heteroaryl ring;each of X¹ and X² is independently selected from N and C(R⁶); Z is

—CD(R⁵)—, or —CH(R⁵)—, wherein “1” represents a point of attachment toN(R⁸); and “2” represents a point of attachment to ring A; each R¹ andeach R⁷ is independently selected from C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₁-C₆ alkoxy, halo, C₁-C₆ heteroalkyl, cycloalkyl, aryl,heteroaryl, aryloxy, aralkyl, heterocyclyl, heterocyclylalkyl, nitro,cyano, —C(O)R^(c), —OC(O)R^(c), —C(O)OR^(d), —(C₁-C₆alkylene)-C(O)R^(c), —SR^(d), —S(O)₂R^(c), —S(O)₂—N(R^(d))(R^(d)),—(C₁-C₆ alkylene)-S(O)₂R^(c), —(C₁-C₆ alkylene)-S(O)₂—N(R^(d))(R^(d)),—N(R^(d))(R^(d)), —C(O)—N(R^(d))(R^(d)), —N(R^(d))—C(O)R^(c),—N(R^(d))—C(O)OR^(c), —(C₁-C₆ alkylene)-N(R^(d))—C(O)R^(c),—N(R^(d))S(O)₂R, and —P(O)(R^(c))(R^(c)); wherein each of alkyl,alkenyl, alkynyl, alkoxy, heteroalkyl, cycloalkyl, aryl, heteroaryl,aryloxy, aralkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a); or two R¹ or two R⁷ are takentogether with the carbon atoms to which they are attached form acycloalkyl or heterocyclyl ring independently substituted with 0-5occurrences of R^(b);each of R², R³ if present, and R⁴ is independently selected fromhydrogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, halo, hydroxyl, cyano, C₁-C₆heteroalkyl, and —N(R^(d))(R^(d)); wherein each of alkyl, alkoxy, andheteroalkyl is optionally and independently substituted with 0-5occurrences of R^(a);each of R⁵ and R⁸ is independently selected from hydrogen, deuterium,C₁-C₆ alkyl, and C₁-C₆ heteroalkyl; wherein each alkyl and heteroalkylis optionally and independently substituted with 0-5 occurrences ofR^(a); each R⁶ is independently selected from hydrogen, C₁-C₆ alkyl,C₁-C₆ alkoxy, halo, cyano, C₁-C₆ heteroalkyl, and —N(R^(d))(R^(d));wherein each alkyl, alkoxy, and heteroalkyl is optionally andindependently substituted with 0-5 occurrences of R^(a);each R^(a) and each R^(b) is independently selected from C₁-C₆ alkyl,halo, hydroxyl, C₁-C₆ heteroalkyl, C₁-C₆ alkoxy, cycloalkyl,heterocyclyl, or cyano, wherein each of alkyl, heteroalkyl, alkoxy,cycloalkyl and heterocyclyl is independently substituted with 0-5occurrences of R′;each R′ is independently selected from C₁-C₆ alkyl, C₁-C₆ heteroalkyl,halo, hydroxyl, cycloalkyl or cyano; or two R′ together with the atom(s)to which they are attached form a cycloalkyl or heterocyclyl ring; eachR^(c) is independently selected from hydrogen, hydroxyl, halo, thiol,C₁-C₆ alkyl, C₁-C₆ thioalkyl, C₁-C₆ alkoxy, C₁-C₆ heteroalkyl,cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, wherein each of alkyl, thioalkyl, alkoxy,heteroalkyl, cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl,and heterocyclylalkyl is independently substituted with 0-5 occurrencesof R^(a), or two R^(c) together with the atom(s) to which they areattached form a cycloalkyl or heterocyclyl ring independentlysubstituted with 0-5 occurrences of R^(b); each R^(d) is independentlyselected from hydrogen, C₁-C₆ alkyl, C₁-C₆ heteroalkyl, cycloalkyl,cycloalkylalkyl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl,wherein each of alkyl, heteroalkyl, cycloalkyl, cycloalkylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a), or two R^(d) together withthe atom(s) to which they are attached form a cycloalkyl or heterocyclylring independently substituted with 0-5 occurrences of R^(b); m is 0, 1,or 2; and n is 0, 1, 2, or 3.

In some embodiments, the compound has the structural formula (Ia):

or a pharmaceutically acceptable salt thereof, wherein R¹ is halo, C₁-C₄alkyl, C₃-C₆ cycloalkyl, or C₁-C₆ alkoxy; and R¹ is substituted with 0-3occurrences of R^(a).

In some embodiments, R¹ is fluoro, —CH₃, —CH₂CH₃, —CHF₂, —OCH₃, orcyclopropyl.

In some embodiments, R² is selected from hydrogen, —C₁-C₄ alkyl, C₁-C₆alkoxy, hydroxyl, and halo; wherein —C₁-C₄ alkyl or C₁-C₆ alkoxy isoptionally substituted (e.g., with 0-3 occurrences of R^(a), e.g.,cyano, halo). In some embodiments, R² is selected from hydrogen, —C₁-C₄alkyl, C₁-C₆ alkoxy, hydroxyl, and fluoro. In some embodiments, R² ishydrogen, fluoro, —CH₃, —CH₂CH₃, —CH₂OH, —CH₂CN, —OCH₂CF₃, —OCH₂CH₂, orOMe.

In some embodiments, R³ if present is hydrogen.

In some embodiments, R⁴ is selected from hydrogen, hydroxyl, halo,cyano, C₁-C₄ alkyl and O—C₁-C₄ alkyl, wherein each alkyl portion of R⁴is substituted with 0-3 occurrences of R^(a). In some embodiments, R⁴ isselected from hydrogen, fluoro, cyano, hydroxyl, —CH₃, —CH₂CN, —CH₂CH₃,—CH₂CH₂OCH₃, —OCH₃, —OCH₂CF₃, and —OCH₂CH₃. In some embodiments, R⁴ isselected from hydrogen, hydroxyl, and —OCH₃.

In some embodiments, Z is selected from

—CH₂—, and —CH(C₁-C₄ alkyl)-, wherein the C₁-C₄ alkyl is substitutedwith 0-3 occurrences of R^(a). In some embodiments, Z is selected from

—CH₂—, and —CH(CH₃)—.

In some embodiments, each R⁶ is independently selected from hydrogen,halo, cyano, and C₁-C₄ alkyl substituted with 0-3 occurrences of R^(a).In some embodiments, each R⁶ is independently selected from hydrogen,fluoro, cyano, —CH₂F and —CH₃.

In some embodiments, R⁸ is selected from hydrogen and —CH₃.

In some embodiments, ring A is selected from phenyl and a 6-memberedmonocyclic heteroaryl comprising at least one nitrogen ring atom. Insome embodiments, ring A is selected from:

In some embodiments, ring A is selected from

n is 1; R⁷ is selected from 1H-pyrazol-1-yl, azetidin-1-yl, andpyrrolidin-1-yl; and R⁷ is substituted with 0-3 occurrences of R^(b).

In some embodiments, R⁷ is selected from 3-fluoroazetidin-1-yl,3,3-difluoropyrrolidin-1-yl, 3-fluoropyrrolidin-1-yl,3-difluoromethyl-1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,4-chloro-1H-pyrazol-1-yl, 3-difluoromethyl-1H-pyrazol-1-yl,4-difluoromethyl-1H-pyrazol-1-yl, 4-cyclopropyl-1H-pyrazol-1-yl,4-fluoro-1H-pyrazol-1-yl, 3,5-bis(difluoromethyl)-1H-pyrazolyl,3-methyl-1H-pyrazol-1-yl, 4-methyl-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyrazol-1-yl. In some embodiments, R⁷is 4-cyclopropyl-1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl, orpyrazol-1-yl.

In some embodiments, ring A is phenyl; n is 0 or 1; and R⁷ if present is—O—C₁-C₄ alkyl.

In some embodiments, n is 0, or n is 1 and R⁷ is selected from —OCH₃ and—OCH₂CH₃.

In another aspect, the disclosure features a compound of Formula (II) ora pharmaceutically acceptable salt thereof, wherein:

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and C(R¹³); each Y¹ and Y² is independently selected from N andCH, wherein no more than one of Y¹ and Y² is N; Q is selected from N, CHand CH₂; R¹¹ is C₁-C₄ alkyl; R¹² is selected from hydrogen and C₁-C₄alkyl; R¹³ if present is selected from hydrogen, cyano and halo; R¹⁴ isselected from hydrogen, halo, cyano, hydroxyl, C₁-C₄ alkyl and C₁-C₄alkoxy; R¹⁵ is selected from hydrogen and C₁-C₄ alkyl; R¹⁶ is selectedfrom hydrogen, and C₁-C₄ alkyl optionally substituted with 1 or moreindependently selected halo; R¹⁷ is selected from hydrogen and C₁-C₄alkyl; each of R^(18a) and R^(19a) if present and R^(18b) and R^(19b) isindependently selected from hydrogen, halo, C₁-C₄ alkyl optionallysubstituted with one or more halo, and C₃-C₆ cycloalkyl; p is 0 or 1;and each

represents a single or a double bond.

In some embodiments, R¹¹ is —CH₃; R¹² is selected from hydrogen and—CH₃; R¹³ if present is selected from hydrogen, cyano and fluoro; R¹⁴ isselected from hydrogen, fluoro, cyano, hydroxyl, —CH₃, —CH₂CH₃, —OCH₃,and —OCH₂CH₃; R¹⁵ is selected from hydrogen and —CH₃; R¹⁶ is selectedfrom hydrogen, —CH₃ and —CHF₂; R¹⁷ is selected from hydrogen and —CH₃;each of R^(18a) and R^(19a) if present is independently selected fromhydrogen and fluoro, wherein at least one of R^(18a) or R^(19a) ishydrogen; each of R^(18b) and R^(19b) is independently selected fromhydrogen, fluoro, chloro, —CH₃, —CHF₂, and cyclopropyl, wherein at leastone of R^(18b) or R^(19b) is hydrogen; and each

is the same.

In some embodiments, p is 1.

In some embodiments, R¹⁴ is selected from hydrogen, fluoro, cyano,hydroxyl, and —OCH₃.

In another aspect, the disclosure features a compound of Formula (III)or a pharmaceutically acceptable salt thereof, wherein:

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and CH; Z′ is selected from

or —CH(R²⁸)—, wherein “1” represents a point of attachment to N(R²⁶);and “2” represents a point of attachment to ring B; ring B is selectedfrom phenyl, pyridinyl, 1H-pyrazolyl, and pyrazinyl; R²¹ is selectedfrom C₃-C₆ cycloalkyl and C₁-C₄ alkyl; R²² is selected from hydrogen andC₁-C₄ alkyl; R²³ is selected from hydrogen and cyano; R²⁴ is selectedfrom hydrogen, hydroxy and halo; R²⁵ is selected from hydrogen, halo,hydroxy, C₁-C₄ alkoxy, —C₁-C₄ alkyl, —C₁-C₄ alkyl-O—C₁-C₄ alkyl, whereineach C₁-C₄ alkyl is optionally substituted with 1 or more substituentsindependently selected from halo and cyano; R²⁶ is selected fromhydrogen and C₁-C₄ alkyl; R²⁷, if present, is independently selectedfrom 1H-pyrazolyl, pyridinyl, and C₁-C₄ alkoxy, wherein the1H-pyrazol-1-yl is optionally substituted with up to 2 substituentsindependently selected from C₁-C₄ alkyl and halo; R²⁸ is selected fromhydrogen and C₁-C₄ alkyl; and o is 0 or 1.

In some embodiments, Z′ is selected from

—CH₂, or —CH(CH₃)—; the portion of the molecule represented by

is selected from

R²¹ is selected from —CH₃ and cyclopropyl; R²² is selected from hydrogenand —CH₃; R²³ is selected from hydrogen and cyano; R²⁴ is selected fromhydrogen, hydroxy and fluoro; R²⁵ is selected from hydrogen, fluoro,hydroxy, —OCH₃, —OCH₂CF₃, —CH₂CH₂OCH₃, —CH₃, —CH₂CH₃, and —CH₂CN; R²⁶ isselected from hydrogen and —CH₃; and R³⁷ is selected from hydrogen,—OCH₃, —OCH₂CH₃, 1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyridin-2-yl. In some embodiments, Z′is —CH₂ or —CH(CH₃)—;

In another aspect, the disclosure features a pharmaceutical compositioncomprising a compound as described herein; and a pharmaceuticallyacceptable carrier.

In another aspect, the disclosure features a method for inhibiting RETactivity in a cell or in a patient, comprising the step of contactingthe cell or administering to the patient a compound as described hereinor a pharmaceutical composition as described herein. In someembodiments, the cell or patient has, or has been identified as having,a RET alteration, e.g., a RET mutation, e.g., a fusion or pointmutation. In some embodiments, the patient comprises a RET-altered cell,cancer, gene, or gene product.

In another aspect, the disclosure features a method for treating asubject suffering from a condition mediated by aberrant RET activity,comprising administering to the subject a therapeutically effectiveamount of a compound as described herein or a pharmaceutical compositionas described herein. In some embodiments, the subject has or has beenidentified as having (e.g., wherein a cancer cell in the subject has orhas been identified as having) a RET alteration, e.g., a RET mutation,e.g., a fusion or point mutation. In some embodiments, a cell, cancer,gene, or gene product from the subject is or has been identified asbeing RET-altered. In some embodiments, the subject has or has beenidentified as having (e.g., a cancer cell in the subject has or has beenidentified as having) a RET alteration, e.g., a RET mutation, e.g., afusion or point mutation.

In some embodiments, the condition mediated by aberrant RET activity isa condition mediated by any RET activity that is not normal e.g., anyactivity due to a RET-altered gene or gene product, which affects theamount or activity of the gene or gene product as compared to the normalor wild-type gene. In some embodiments, the condition mediated byaberrant RET activity is a familial or sporadic cancer, e.g., a solidtumor such as thyroid, lung, breast, or pancreatic. In some embodiments,the condition mediated by aberrant RET activity is irritable bowelsyndrome (IBS). In some embodiments, the aberrant RET activity promotesthe condition, such that inhibition of RET ameliorates at least onesymptom of the condition. In some embodiments, the aberrant RET activitycomprises increased RET activity or expression level, gain of functionmutation, and/or constitutive activation of RET. In some embodiments,the aberrant RET activity corresponds to aberrant amounts of RET, e.g.,aberrant nucleic acid or protein amounts.

In another aspect, the disclosure features a method for treating asubject who has developed resistance to a treatment for a conditionmediated by aberrant RET activity, comprising administering to thesubject a therapeutically effective amount of a compound as describedherein or a pharmaceutical composition as described herein. In someembodiments, the subject has developed resistance to a wild-type RETinhibitor. In some embodiments, the cancer treatment to which thesubject is resistant is a wild-type RET inhibitor that is active againstthe wild-type RET, but less active, e.g., much less active, against oneor more mutated forms of RET. In some embodiments, the wild-type RETinhibitor is selected from ponatinib, cabozanitib, and vandetanib.

In another aspect, the present disclosure provides a use of a compoundor pharmaceutical composition described herein, e.g., a compound ofstructural Formula (I), (Ia), (II), or (III) described herein (e.g., acompound in Table 1) or a pharmaceutically acceptable salt orcomposition thereof, in the manufacture of a medicament for inhibitingRET activity in a cell or in a subject. In some embodiments, the cell orsubject has, or has been identified as having, a RET alteration, e.g., aRET mutation, e.g., a fusion or point mutation. In some embodiments, acell, cancer, gene, or gene product from the subject is or has beenidentified as being RET-altered.

In another aspect, the present disclosure provides a use of a compoundor pharmaceutical composition described herein, e.g., a compound ofstructural Formula (I), (Ia), (II), or (III) described herein (e.g., acompound in Table 1) or a pharmaceutically acceptable salt orcomposition thereof, in the manufacture of a medicament for treating asubject suffering from a condition mediated by aberrant RET activity. Insome embodiments, the subject has, or has been identified as having(e.g., wherein a cancer cell in the subject has, or has been identifiedas having) a RET alteration, e.g., a RET mutation, e.g., a fusion orpoint mutation. In some embodiments, a cell, cancer, gene, or geneproduct from the subject is or has been identified as being RET-altered.

In another aspect, the present disclosure provides a use of a compoundor pharmaceutical composition described herein, e.g., a compound ofstructural Formula (I), (Ia), (II), or (III) described herein (e.g., acompound in Table 1) or a pharmaceutically acceptable salt orcomposition thereof, in the manufacture of a medicament for treating asubject who has developed resistance to a cancer treatment.

In another aspect, the present disclosure provides a method ofpreventing development of one or more RET-altered cell, cancer, gene, orgene product, in a cell or in a patient, comprising the step ofcontacting the cell or administering to the patient a compound accordingto structural Formula (I), (Ia), (II), or (III).

In another aspect, the present disclosure provides a method of treatinga patient suffering from cancer comprising administering to the patienta compound described herein (e.g., a compound having a lower IC₅₀ forRET than for KDR, a compound having a similar IC₅₀ for wild-type RETcompared to mutant RET, and/or a compound of structural Formula (I),(Ia), (II), or (III) described herein (e.g., a compound in Table 1)), ora pharmaceutical composition comprising the compound, wherein thesubject has a RET-altered cell, cancer, gene, or gene product that isresponsive to the compound. In some embodiments, the disclosure providesa method of treating cancer in a patient, said method comprisingadministering an effective amount of a compound described herein to apatient having a RET-altered cell, cancer, gene, or gene product that isresponsive to the compound.

In another aspect, the present disclosure provides a method of treatinga subject suffering from cancer comprising the steps of:

-   -   a. receiving information related to a RET sequence, e.g.,        information related to a RET-altered gene or gene product, e.g.,        having a RET fusion or point mutation; and    -   b. administering to the subject a compound described herein        (e.g., a compound having a lower IC₅₀ for RET than for KDR, a        compound having a similar IC₅₀ for wild-type RET compared to        mutant RET, and/or a compound of structural Formula (I), (Ia),        (II), or (III) described herein (e.g., a compound in Table 1)),        or a pharmaceutical composition comprising the compound, if the        information indicates a RET-altered cell, cancer, gene, or gene        product.

In some embodiments, the subject is administered a cancer treatmentother than a compound of structural Formula (I), (Ia), (II), or (III) ifthe information indicates that the subject has a wild-type RET sequence.In some embodiments, the cancer treatment is a wild-type RET inhibitor.

In another aspect, the present disclosure provides a method of treatingcancer in a subject, said method comprising:

a. obtaining a biological sample (e.g., a tumor biopsy) from a humansubject;

b. detecting whether a RET-altered cell, cancer, gene, or gene product,e.g., having a fusion or point mutation, is present in the biologicalsample;

c. identifying the subject as responsive to a compound described herein(e.g., a compound having a lower IC₅₀ for RET than for KDR, a compoundhaving a similar IC₅₀ for wild-type RET compared to mutant RET, and/or acompound of structural Formula (I), (Ia), (II), or (III) describedherein (e.g., a compound in Table 1)) when the presence of theRET-altered cell, cancer, gene, or gene product in the biological sampleis detected; and

d. administering an effective amount of the compound to the subject.

In another aspect, the present disclosure provides a method of treatingcancer in a subject, said method comprising:

-   -   a. determining if, having determined if, or receiving        information that the subject has a RET-altered cell, cancer,        gene, or gene product, e.g., having a fusion or point mutation;    -   b. identifying the subject as responsive to a compound described        herein (e.g., a compound having a lower IC₅₀ for RET than for        KDR, a compound having a similar IC₅₀ for wild-type RET compared        to mutant RET, and/or a compound of structural Formula (I),        (Ia), (II), or (III) described herein (e.g., a compound in Table        1)) when the subject has a RET-altered cell, cancer, gene, or        gene product; and    -   c. administering an effective amount of the compound to the        subject.

In another aspect, the present disclosure provides a method ofdiagnosing cancer in a subject, said method comprising:

a. obtaining a biological sample (e.g., a tumor biopsy) from a humansubject;

b. detecting whether a RET-altered cell, cancer, gene, or gene product,e.g., having a fusion or point mutation, is present in the biologicalsample;

c. diagnosing the subject with cancer when the presence of theRET-altered cell, cancer, gene, or gene product in the biological sampleis detected.

In another aspect, the present disclosure provides a method ofpredicting the efficacy of a compound described herein (a compoundhaving a lower IC₅₀ for RET than for KDR, a compound having a similarIC₅₀ for wild-type RET compared to mutant RET, and/or a compound ofstructural Formula (I), (Ia), (II), or (III) described herein (e.g., acompound in Table 1)) in a treatment of cancer in a subject comprisingthe step of:

determining if, having determined if, or receiving information that thesubject has a RET-altered cell, cancer, gene, or gene product, e.g.,having a mutation, e.g., a fusion or point mutation, e.g., by a methodselected from hybridization-based methods, amplification-based methods,microarray analysis, flow cytometry analysis, DNA sequencing,next-generation sequencing (NGS), primer extension, PCR, in situhybridization, dot blot, and Southern blot;

wherein said determining if, having determined if, or receivinginformation is predictive of efficacy of the compound in the treatment.

In some embodiments of any of the methods and uses herein, the methodfurther comprises administering to the subject a compound describedherein (e.g., a compound having a lower IC₅₀ for RET than for KDR, acompound having a similar IC₅₀ for wild-type RET compared to mutant RET,and/or a compound of structural Formula (I), (Ia), (II), or (III)described herein (e.g., a compound in Table 1)), e.g., responsive to adetermination or diagnosis made by a method described herein.

In some embodiments of any of the methods and uses herein, theRET-altered cell, cancer, gene, or gene product comprises a fusionmutation, e.g., CCDC6-RET or KIF5B-RET or a fusion of Table 3. In someembodiments, the RET-altered cell, cancer, gene, or gene productcomprises a point mutation of amino acid position 634, 918, or 804 ofRET, or a point mutation at a position listed in Table 4, e.g., a pointmutation specified in Table 4. In some embodiments, the RET-alteredcell, cancer, gene, or gene product comprises a point mutation at aminoacid 634, 918, or 804, 806, 810, 865, 870, 891, e.g., is selected fromRET C634W, M918T, V804L, V804E, V804M, V804L, V806C, Y806C, Y806S,Y806N, Y806H, G810R, G810S, L865V, L870F, and S891A. In someembodiments, the RET alteration is located at least partially within, oris located wholly within one or more of: the N-terminal extracellulardomain (e.g., within one or more cadherin-like repeats and/or thecysteine-rich region), the transmembrane domain, or the tyrosine kinasedomain (e.g., within one or more of the ATP binding site or protonacceptor site). In some embodiments, the RET fusion further comprises apoint mutation, e.g., KIF5B-RET (V804L) or KIF5B-RET (V804M).

In some embodiments of any of the methods and uses herein, the subjectsuffers from a cancer selected from colorectal cancer, lung cancer(e.g., a lung adenocarcinoma, e.g., NSCLC), thyroid cancer (e.g.,medullary thyroid cancer), or leukemia. In some embodiments, the subjectsuffers from a cancer listed herein, e.g., in Table 3.

In some embodiments, the cancer is wild-type RET.

In some embodiments, the cancer is lung cancer, e.g., a lungadenocarcinoma, and the RET-altered cell, cancer, gene, or gene productcomprises a CCDC6-RET fusion. In some embodiments, the cancer is lungadenocarcinoma and the RET-altered cell, cancer, gene, or gene productcomprises a KIF5B-RET fusion. In some embodiments, the cancer is lungadenocarcinoma and the RET-altered cell, cancer, gene, or gene productcomprises KIF5B-RET (V804L). In some embodiments, the lungadenocarcinoma is NSCLC.

In some embodiments, the cancer is thyroid cancer, e.g., medullarythyroid cancer, and the RET-altered cell, cancer, gene, or gene productcomprises a C634W mutation.

In some embodiments, the cancer is leukemia and the RET-altered cell,cancer, gene, or gene product comprises a KIF5B-RET fusion.

In some embodiments, the cancer is thyroid cancer and the RET-alteredcell, cancer, gene, or gene product comprises a M918T mutation. In someembodiments, the cancer is medullary thyroid cancer and the RET-alteredcell, cancer, gene, or gene product comprises a M918T mutation.

In some embodiments, the cancer is leukemia and the RET-altered cell,cancer, gene, or gene product comprises comprises KIF5B-RET (V804L) orKIF5B-RET (V804M) fusion.

In some embodiments, the cancer is thyroid cancer and the RET-alteredcell, cancer, gene, or gene product comprises comprises a CCDC6-RETfusion. In some embodiments, the cancer is colorectal cancer and theRET-altered cell, cancer, gene, or gene product comprises CCDC6-RETfusion. In some embodiments, the cancer is colorectal cancer and theRET-altered cell, cancer, gene, or gene product comprises CCDC6-RET(V804M) fusion.

In some embodiments of any of the methods and uses herein, the compoundor pharmaceutical composition has a lower IC₅₀ for RET than for KDR,e.g., has a KDR/RET IC₅₀ ratio of at least 3×, 4×, 5×, 10×, 20×, 50×, or100×, and optionally up to 50× or 100×. In some embodiments, thecompound or pharmaceutical composition has a KDR/RET IC₅₀ ratio ofbetween 3×-4×, 4×-5×, 5×-100×, 100×-20×, 20×-50×, or 50×-100×. In someembodiments, the compound or pharmaceutical composition has a similarIC₅₀ for wild-type RET compared to mutant RET (e.g., for V804L RET orV804E RET), e.g., has a wild-type/mutant IC₅₀ ratio of no more than 3×,2×, 1.5×, 1×, or 0.5×, e.g., of between 3× and 0.5×.

In some embodiments of any of the methods and uses herein, the subjectdoes not develop a RET-altered cell, cancer, gene, or gene product forat least 1, 2, 3, 6, 9, 12, 24, or 36 months after initiation ofadministration of the compound. In some embodiments, the compound isadministered as a first line therapy. In some embodiments, the compoundis administered to a treatment-naïve subject. In some embodiments, thecompound is not administered in combination with another kinaseinhibitor. In some embodiments, the compound is not administered incombination with another RET inhibitor. For example, the compound can beadministered as a monotherapy or in combination with one or other agentswhich are not kinase inhibitors, e.g., not RET inhibitors.

In some embodiments of any of the methods and uses herein, the compoundis administered at an amount sufficient to reach at least 70%, 80%, 90%,or 95% inhibition of RET in vivo.

EMBODIMENTS OF THE DISCLOSURE Definitions

As used herein, the terms a “patient,” “subject,” “individual,” and“host” refer to either a human or a non-human animal suffering from orsuspected of suffering from a disease or disorder associated withaberrant RET expression (i.e., increased RET activity caused bysignaling through RET) or biological activity.

“Treat” and “treating” such a disease or disorder refers to amelioratingat least one symptom of the disease or disorder. These terms, when usedin connection with a condition such as a cancer, refer to one or moreof: impeding growth of the cancer, causing the cancer to shrink byweight or volume, extending the expected survival time of the patient,inhibiting tumor growth, reducing tumor mass, reducing size or number ofmetastatic lesions, inhibiting the development of new metastaticlesions, prolonging survival, prolonging progression-free survival,prolonging time to progression, and/or enhancing quality of life.

The term “preventing” when used in relation to a condition or diseasesuch as cancer, refers to a reduction in the frequency of, or delay inthe onset of, symptoms of the condition or disease. Thus, prevention ofcancer includes, for example, reducing the number of detectablecancerous growths in a population of patients receiving a prophylactictreatment relative to an untreated control population, and/or delayingthe appearance of detectable cancerous growths in a treated populationversus an untreated control population, e.g., by a statistically and/orclinically significant amount.

The term “therapeutic effect” refers to a beneficial local or systemiceffect in animals, particularly mammals, and more particularly humans,caused by administration of a compound or composition of the disclosure.The phrase “therapeutically-effective amount” means that amount of acompound or composition of the disclosure that is effective to treat adisease or condition caused by over expression of RET or aberrant RETbiological activity at a reasonable benefit/risk ratio. Thetherapeutically effective amount of such substance will vary dependingupon the subject and disease or condition being treated, the weight andage of the subject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofskill in the art.

As used herein, “developing resistance” means that when a drug is firstadministered to the patient, the patient's symptoms improve, whethermeasured by decrease in tumor volume, a decrease in the number of newlesions, or some other means that a physician uses to judge diseaseprogression, however, those symptoms stop improving, or even worsen atsome point. At that time, the patient is said to have developedresistance to the drug.

“Alteration” as used herein, of a gene or gene product (e.g., the RETgene or gene product) refers to the presence of a mutation or mutationswithin the gene or gene product, e.g., a mutation, which of the gene orgene product, as compared to the normal or wild-type gene. Thealteration can be in amount, structure, and/or activity in a cancertissue or cancer cell, as compared to its amount, structure, and/oractivity, in a normal or healthy tissue or cell (e.g., a control), andis associated with a disease or condition, such as cancer. For example,an alteration which is associated with cancer, or predictive ofresponsiveness to an anti-cancer therapeutic, can have an alterednucleotide sequence (e.g., a mutation), amino acid sequence, chromosomaltranslocation, intra-chromosomal inversion, copy number, expressionlevel, protein level, protein activity, or methylation status, in acancer tissue or cancer cell, as compared to a normal, healthy tissue orcell. Exemplary mutations include, but are not limited to, pointmutations (e.g., silent, missense, or nonsense), deletions, insertions,inversions, linking mutations, duplications, translocations, inter- andintra-chromosomal rearrangements. Mutations can be present in the codingor non-coding region of the gene, e.g., a 3′ UTR or 5′ UTR.

A subject having “altered RET” refers to a subject comprising a RETalteration, e.g., in one or more of their cancer cells.

A “RET-altered” cell, cancer, gene, or gene product refers to a cell,cancer, gene, or gene product comprising a RET alteration as describedherein.

“Aliphatic group” means a straight-chain, branched-chain, or cyclichydrocarbon group and includes saturated and unsaturated groups, such asan alkyl group, an alkenyl group, and an alkynyl group.

“Alkylene” refers to a divalent radical of an alkyl group, e.g., —CH₂—,—CH₂CH₂—, and —CH₂CH₂CH₂—.

“Alkenyl” means an aliphatic group containing at least one double bond.

“Alkoxyl” or “alkoxy” means an alkyl group having an oxygen radicalattached thereto. Representative alkoxyl groups include methoxy, ethoxy,propyloxy, tert-butoxy and the like. The term “haloalkoxy” refers to analkoxy in which one or more hydrogen atoms are replaced by halo, andincludes alkoxy moieties in which all hydrogens have been replaced byhalo (e.g., perfluoroalkoxy).

“Alkyl” refers to a monovalent radical of a saturated straight orbranched hydrocarbon, such as a straight or branched group of 1-12,1-10, or 1-6 carbon atoms, referred to herein as C₁-C₁₂ alkyl, C₁-C₁₀alkyl, and C₁-C₆ alkyl, respectively. Exemplary alkyl groups include,but are not limited to, methyl, ethyl, propyl, isopropyl,2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl,3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl,2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl,2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl,2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl,isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl,etc.

“Alkenylene” refers to an alkenyl group having two connecting points.For example, “ethenylene” represents the group —CH═CH—. Alkenylenegroups can also be in an unsubstituted form or substituted form with oneor more substituents.

“Alkynyl” refers to a straight or branched hydrocarbon chain containing2-12 carbon atoms and characterized in having one or more triple bonds.Examples of alkynyl groups include, but are not limited to, ethynyl,propargyl, and 3-hexynyl. One of the triple bond carbons may optionallybe the point of attachment of the alkynyl substituent.

“Alkynylene” refers to an alkynyl having two connecting points. Forexample, “ethynylene” represents the group —C≡C—. Alkynylene groups canalso be in an unsubstituted form or substituted form with one or moresubstituents.

“Hydroxyalkylene” or “hydroxyalkyl” refers to an alkylene or alkylmoiety in which an alkylene or alkyl hydrogen atom is replaced by ahydroxyl group. Hydroxyalkylene or hydroxyalkyl includes groups in whichmore than one hydrogen atom has been replaced by a hydroxyl group.

“Aromatic ring system” is art-recognized and refers to a monocyclic,bicyclic or polycyclic hydrocarbon ring system, wherein at least onering is aromatic.

“Aryl” refers to a monovalent radical of an aromatic ring system.Representative aryl groups include fully aromatic ring systems, such asphenyl, naphthyl, and anthracenyl, and ring systems where an aromaticcarbon ring is fused to one or more non-aromatic carbon rings, such asindanyl, phthalimidyl, naphthimidyl, or tetrahydronaphthyl, and thelike.

“Arylalkyl” or “aralkyl” refers to an alkyl moiety in which an alkylhydrogen atom is replaced by an aryl group. Aralkyl includes groups inwhich more than one hydrogen atom has been replaced by an aryl group.Examples of“arylalkyl” or “aralkyl” include benzyl, 2-phenylethyl,3-phenylpropyl, 9-fluorenyl, benzhydryl, and trityl groups.

“Aryloxy” refers to —O-(aryl), wherein the aryl moiety is as definedherein.

“Halo” refers to a radical of any halogen, e.g., —F, —Cl, —Br, or —I.

“Haloalkyl” and “haloalkoxy” refers to alkyl and alkoxy structures thatare substituted with one or more halo groups or with combinationsthereof. For example, the terms “fluoroalkyl” and “fluoroalkoxy” includehaloalkyl and haloalkoxy groups, respectively, in which the halo isfluorine. “Haloalkylene” refers to a divalent alkyl, e.g., —CH₂—,—CH₂CH₂—, and —CH₂CH₂CH₂—, in which one or more hydrogen atoms arereplaced by halo, and includes alkyl moieties in which all hydrogenshave been replaced by halo.

“Heteroalkyl” refers to an optionally substituted alkyl, which has oneor more skeletal chain atoms selected from an atom other than carbon,e.g., oxygen, nitrogen, sulfur, phosphorus or combinations thereof. Anumerical range may be given, e.g. C₁-C₆ heteroalkyl which refers to thenumber of carbons in the chain, which in this example includes 1 to 6carbon atoms. For example, a —CH₂OCH₂CH₃ radical is referred to as a“C₃” heteroalkyl. Connection to the rest of the molecule may be througheither a heteroatom or a carbon in the heteroalkyl chain.“Heteroalkylene” refers to a divalent optionally substituted alkyl,which has one or more skeletal chain atoms selected from an atom otherthan carbon, e.g., oxygen, nitrogen, sulfur, phosphorus or combinationsthereof.

“Carbocyclic ring system” refers to a monocyclic, bicyclic or polycyclichydrocarbon ring system, wherein each ring is either completelysaturated or contains one or more units of unsaturation, but where noring is aromatic.

“Carbocyclyl” refers to a monovalent radical of a carbocyclic ringsystem. Representative carbocyclyl groups include cycloalkyl groups(e.g., cyclobutyl, cyclopentyl, cyclohexyl and the like), andcycloalkenyl groups (e.g., cyclopentenyl, cyclohexenyl,cyclopentadienyl, and the like).

“Cycloalkyl” refers to a cyclic, bicyclic, tricyclic, or polycyclicnon-aromatic hydrocarbon groups having 3 to 12 carbons. Anysubstitutable ring atom can be substituted (e.g., by one or moresubstituents). The cycloalkyl groups can contain fused or spiro rings.Fused rings are rings that share a common carbon atom. Examples ofcycloalkyl moieties include, but are not limited to, cyclopropyl,cyclohexyl, methylcyclohexyl, adamantyl, and norbornyl.

“Cycloalkylalkyl” refers to a -(cycloalkyl)-alkyl radical wherecycloalkyl and alkyl are as disclosed herein. The “cycloalkylalkyl” isbonded to the parent molecular structure through the alkyl group.

“Heteroaromatic ring system” is art-recognized and refers to monocyclic,bicyclic or polycyclic ring system wherein at least one ring is botharomatic and comprises at least one heteroatom (e.g., N, O or S); andwherein no other rings are heterocyclyl (as defined below). In certaininstances, a ring which is aromatic and comprises a heteroatom contains1, 2, 3, or 4 ring heteroatoms in such ring.

“Heteroaryl” refers to a monovalent radical of a heteroaromatic ringsystem. Representative heteroaryl groups include ring systems where (i)each ring comprises a heteroatom and is aromatic, e.g., imidazolyl,oxazolyl, thiazolyl, triazolyl, pyrrolyl, furanyl, thiophenyl pyrazolyl,pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, indolizinyl, purinyl,naphthyridinyl, and pteridinyl; (ii) each ring is aromatic orcarbocyclyl, at least one aromatic ring comprises a heteroatom and atleast one other ring is a hydrocarbon ring or e.g., indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, carbazolyl, acridinyl, phenazinyl,phenothiazinyl, phenoxazinyl, pyrido[2,3-b]-1,4-oxazin-3-(4H)-one,5,6,7,8-tetrahydroquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl; and(iii) each ring is aromatic or carbocyclyl, and at least one aromaticring shares a bridgehead heteroatom with another aromatic ring, e.g.,4H-quinolizinyl.

“Heterocyclic ring system” refers to monocyclic, bicyclic and polycyclicring systems where at least one ring is saturated or partiallyunsaturated (but not aromatic) and comprises at least one heteroatom. Aheterocyclic ring system can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted.

“Heterocyclyl” refers to a monovalent radical of a heterocyclic ringsystem. Representative heterocyclyls include ring systems in which (i)every ring is non-aromatic and at least one ring comprises a heteroatom,e.g., tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl,pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl,diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl;(ii) at least one ring is non-aromatic and comprises a heteroatom and atleast one other ring is an aromatic carbon ring, e.g.,1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl; and (iii)at least one ring is non-aromatic and comprises a heteroatom and atleast one other ring is aromatic and comprises a heteroatom, e.g.,3,4-dihydro-1H-pyrano[4,3-c]pyridine, and1,2,3,4-tetrahydro-2,6-naphthyridine. In some embodiments, heterocyclylcan include:

wherein the point of attachment to the base structure can be through anyof the atoms on the heterocyclyl, e.g., through a carbon atom or anitrogen atom of the heterocyclyl.

“Heterocyclylalkyl” refers to an alkyl group substituted with aheterocyclyl group. The “heterocyclylalkyl” is bonded to the parentmolecular structure through the alkyl group.

“Cyano” refers to a —CN radical.

“Nitro” refers to —NO₂.

“Hydroxy” or “hydroxyl” refers to —OH.

“Hydroxyalkylene” refers to a divalent alkyl, e.g., —CH₂—, —CH₂CH₂—, and—CH₂CH₂CH₂—, in which one or more hydrogen atoms are replaced by ahydroxy, and includes alkyl moieties in which all hydrogens have beenreplaced by hydroxy.

“Substituted”, whether preceded by the term “optionally” or not, meansthat one or more hydrogens of the designated moiety are replaced with asuitable substituent. Unless otherwise indicated, an “optionallysubstituted” group may have a suitable substituent at each substitutableposition of the group, and when more than one position in any givenstructure may be substituted with more than one substituent selectedfrom a specified group, the substituent may be either the same ordifferent at each position. Combinations of substituents envisionedunder this disclosure are preferably those that result in the formationof stable or chemically feasible compounds. The term “stable”, as usedherein, refers to compounds that are not substantially altered whensubjected to conditions to allow for their production, detection, and,in certain embodiments, their recovery, purification, and use for one ormore of the purposes disclosed herein.

As used herein, the definition of each expression, e.g., alkyl, m, n,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

Certain compounds of the present disclosure may exist in particulargeometric or stereoisomeric forms. The present disclosure contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the disclosure. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in thisdisclosure.

If, for instance, a particular enantiomer of compound of the presentdisclosure is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

Unless otherwise indicated when a disclosed compound is named ordepicted by a structure without specifying the stereochemistry and hasone or more chiral centers, it is understood to represent all possiblestereoisomers of the compound, as well as enantiomeric mixtures thereof.

The “enantiomeric excess” or “% enantiomeric excess” of a compositioncan be calculated using the equation shown below. In the example shownbelow a composition contains 90% of one enantiomer, e.g., the Senantiomer, and 10% of the other enantiomer, i.e., the R enantiomer.

ee=(90−10)/100=80%.

Thus, a composition containing 90% of one enantiomer and 10% of theother enantiomer is said to have an enantiomeric excess of 80%.

The compounds or compositions described herein may contain anenantiomeric excess of at least 50%, 75%, 90%, 95%, or 99% of one formof the compound, e.g., the S-enantiomer. In other words such compoundsor compositions contain an enantiomeric excess of the S enantiomer overthe R enantiomer.

The compounds described herein may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example deuterium (²H), tritium (³H),carbon-13 (¹³C), or carbon-14 (¹⁴C). All isotopic variations of thecompounds disclosed herein, whether radioactive or not, are intended tobe encompassed within the scope of the present disclosure. In addition,all tautomeric forms of the compounds described herein are intended tobe within the scope of the disclosure.

The compound can be useful as the free base or as a salt. Representativesalts include the hydrobromide, hydrochloride, sulfate, bisulfate,phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate,laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate,fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate,lactobionate, and laurylsulphonate salts and the like. (See, forexample, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci.66:1-19.)

Compounds

In one aspect, the disclosure features a compound of Formula (I) or apharmaceutically acceptable salt thereof, wherein:

ring A is an aryl or heteroaryl ring;each of X¹ and X² is independently selected from N and C(R⁶); Z is

—CD(R⁵)—, or —CH(R⁵)—, wherein “1” represents a point of attachment toN(R⁸); and “2” represents a point of attachment to ring A; each R¹ andeach R⁷ is independently selected from C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₁-C₆ alkoxy, halo, C₁-C₆ heteroalkyl, cycloalkyl, aryl,heteroaryl, aryloxy, aralkyl, heterocyclyl, heterocyclylalkyl, nitro,cyano, —C(O)R^(c), —OC(O)R^(c), —C(O)OR^(d), —(C₁-C₆alkylene)-C(O)R^(c), —SR^(d), —S(O)₂R^(c), —S(O)₂—N(R^(d))(R^(d)),—(C₁-C₆ alkylene)-S(O)₂R^(c), —(C₁-C₆ alkylene)-S(O)₂—N(R^(d))(R^(d)).N(R^(d))(R^(d)), —C(O)—N(R^(d))(R^(d)), —N(R^(d))—C(O)R^(c),—N(R^(d))—C(O)OR^(c), —(C₁-C₆ alkylene)-N(R^(d))—C(O)R^(c),—N(R^(d))S(O)₂R^(c), and —P(O)(R^(c))(R^(c)); wherein each of alkyl,alkenyl, alkynyl, alkoxy, heteroalkyl, cycloalkyl, aryl, heteroaryl,aryloxy, aralkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a); or two R¹ or two R⁷ are takentogether with the carbon atoms to which they are attached form acycloalkyl or heterocyclyl ring independently substituted with 0-5occurrences of R^(b);each of R², R³ if present, and R⁴ is independently selected fromhydrogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, halo, hydroxyl, cyano, C₁-C₆heteroalkyl, and —N(R^(d))(R^(d)); wherein each of alkyl, alkoxy, andheteroalkyl is optionally and independently substituted with 0-5occurrences of R^(a);each of R⁵ and R⁸ is independently selected from hydrogen, deuterium,C₁-C₆ alkyl, and C₁-C₆ heteroalkyl; wherein each alkyl and heteroalkylis optionally and independently substituted with 0-5 occurrences ofR^(a); each R⁶ is independently selected from hydrogen, C₁-C₆ alkyl,C₁-C₆ alkoxy, halo, cyano, C₁-C₆ heteroalkyl, and —N(R^(d))(R^(d));wherein each alkyl, alkoxy, and heteroalkyl is optionally andindependently substituted with 0-5 occurrences of R^(a);each R^(a) and each R^(b) is independently selected from C₁-C₆ alkyl,halo, hydroxyl, C₁-C₆ heteroalkyl, C₁-C₆ alkoxy, cycloalkyl,heterocyclyl, or cyano, wherein each of alkyl, heteroalkyl, alkoxy,cycloalkyl and heterocyclyl is independently substituted with 0-5occurrences of R′;each R′ is independently selected from C₁-C₆ alkyl, C₁-C₆ heteroalkyl,halo, hydroxyl, cycloalkyl or cyano; or two R′ together with the atom(s)to which they are attached form a cycloalkyl or heterocyclyl ring; eachR^(c) is independently selected from hydrogen, hydroxyl, halo, thiol,C₁-C₆ alkyl, C₁-C₆ thioalkyl, C₁-C₆ alkoxy, C₁-C₆ heteroalkyl,cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, wherein each of alkyl, thioalkyl, alkoxy,heteroalkyl, cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl,and heterocyclylalkyl is independently substituted with 0-5 occurrencesof R^(a), or two R^(c) together with the atom(s) to which they areattached form a cycloalkyl or heterocyclyl ring independentlysubstituted with 0-5 occurrences of R^(b); each R^(d) is independentlyselected from hydrogen, C₁-C₆ alkyl, C₁-C₆ heteroalkyl, cycloalkyl,cycloalkylalkyl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl,wherein each of alkyl, heteroalkyl, cycloalkyl, cycloalkylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a), or two R^(d) together withthe atom(s) to which they are attached form a cycloalkyl or heterocyclylring independently substituted with 0-5 occurrences of R^(b); m is 0, 1,or 2; and n is 0, 1, 2, or 3.

In some embodiments, the compound has the structural formula (Ia):

or a pharmaceutically acceptable salt thereof, wherein R¹ is halo, C₁-C₄alkyl, C₃-C₆ cycloalkyl, or C₁-C₆ alkoxy; and R¹ is substituted with 0-3occurrences of R^(a).

In some embodiments, R¹ is fluoro, —CH₃—CH₂CH₃, —CHF₂, —OCH₃, orcyclopropyl.

In some embodiments, R² is selected from hydrogen, —C₁-C₄ alkyl, C₁-C₆alkoxy, hydroxyl, and halo; wherein —C₁-C₄ alkyl or C₁-C₆ alkoxy isoptionally substituted (e.g., with 0-3 occurrences of R^(a), e.g.,cyano, halo). In some embodiments, R² is selected from hydrogen, —C₁-C₄alkyl, C₁-C₆ alkoxy, hydroxyl, and fluoro. In some embodiments, R² ishydrogen, fluoro, —CH₃, —CH₂CH₃, —CH₂OH, —CH₂CN, —OCH₂CF₃, —OCH₂CH₂, orOMe.

In some embodiments, R³ if present is hydrogen.

In some embodiments, R⁴ is selected from hydrogen, hydroxyl, halo,cyano, C₁-C₄ alkyl and O—C₁-C₄ alkyl, wherein each alkyl portion of R⁴is substituted with 0-3 occurrences of R^(a).

In some embodiments, R⁴ is selected from hydrogen, fluoro, cyano,hydroxyl, —CH₃, —CH₂CN, —CH₂CH₃, —CH₂CH₂OCH₃, —OCH₃, —OCH₂CF₃, and—OCH₂CH₃. In some embodiments, R⁴ is selected from hydrogen, hydroxyl,and —OCH₃.

In some embodiments, Z is selected from

—CH₂—, and —CH(C₁-C₄ alkyl)-, wherein the C₁-C₄ alkyl is substitutedwith 0-3 occurrences of R^(a). In some embodiments, Z is selected from

—CH₂—, and —CH(CH₃)—.

In some embodiments, each R⁶ is independently selected from hydrogen,halo, cyano, and C₁-C₄ alkyl substituted with 0-3 occurrences of R^(a).In some embodiments, each R⁶ is independently selected from hydrogen,fluoro, cyano, —CH₂F and —CH₃.

In some embodiments, R⁸ is selected from hydrogen and —CH₃.

In some embodiments, ring A is selected from phenyl and a 6-memberedmonocyclic heteroaryl comprising at least one nitrogen ring atom. Insome embodiments, ring A is selected from:

In some embodiments, ring A is selected from

n is 1; R⁷ is selected from 1H-pyrazol-1-yl, azetidin-1-yl, andpyrrolidin-1-yl; and R⁷ is substituted with 0-3 occurrences of R^(b).

In some embodiments, R⁷ is selected from 3-fluoroazetidin-1-yl,3,3-difluoropyrrolidin-1-yl, 3-fluoropyrrolidin-1-yl,3-difluoromethyl-1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,4-chloro-1H-pyrazol-1-yl, 3-difluoromethyl-1H-pyrazol-1-yl,4-difluoromethyl-1H-pyrazol-1-yl, 4-cyclopropyl-1H-pyrazol-1-yl,4-fluoro-1H-pyrazol-1-yl, 3,5-bis(difluoromethyl)-1H-pyrazolyl,3-methyl-1H-pyrazol-1-yl, 4-methyl-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyrazol-1-yl. In some embodiments, R⁷is 4-cyclopropyl-1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl, orpyrazol-1-yl.

In some embodiments, ring A is phenyl; n is 0 or 1; and R⁷ if present is—O—C₁-C₄ alkyl.

In some embodiments, n is 0, or n is 1 and R⁷ is selected from —OCH₃ and—OCH₂CH₃.

In another aspect, the disclosure features a compound of Formula (II) ora pharmaceutically acceptable salt thereof, wherein:

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and C(R¹³); each Y¹ and Y² is independently selected from N andCH, wherein no more than one of Y¹ and Y² is N; Q is selected from N, CHand CH₂; R¹¹ is C₁-C₄ alkyl; R¹² is selected from hydrogen and C₁-C₄alkyl; R¹³ if present is selected from hydrogen, cyano and halo; R¹⁴ isselected from hydrogen, halo, cyano, hydroxyl, C₁-C₄ alkyl and C₁-C₄alkoxy; R¹⁵ is selected from hydrogen and C₁-C₄ alkyl; R¹⁶ is selectedfrom hydrogen, and C₁-C₄ alkyl optionally substituted with 1 or moreindependently selected halo; R¹⁷ is selected from hydrogen and C₁-C₄alkyl; each of R^(18a) and R^(19a) if present and R^(18b) and R^(19b) isindependently selected from hydrogen, halo. C₁-C₄ alkyl optionallysubstituted with one or more halo, and C₃-C₆ cycloalkyl; p is 0 or 1;and each

represents a single or a double bond.

In some embodiments, R¹¹ is —CH₃; R¹² is selected from hydrogen and—CH₃; R¹³ if present is selected from hydrogen, cyano and fluoro; R¹⁴ isselected from hydrogen, fluoro, cyano, hydroxyl, —CH₃, —CH₂CH₃, —OCH₃,and —OCH₂CH₃; R¹⁵ is selected from hydrogen and —CH₃; R¹⁶ is selectedfrom hydrogen, —CH₃ and —CHF₂; R¹⁷ is selected from hydrogen and —CH₃;each of R^(18a) and R^(19a) if present is independently selected fromhydrogen and fluoro, wherein at least one of R^(18a) or R^(19a) ishydrogen; each of R^(18b) and R^(19b) is independently selected fromhydrogen, fluoro, chloro, —CH₃, —CHF₂, and cyclopropyl, wherein at leastone of R^(18b) or R^(19b) is hydrogen; and each

is the same.

In some embodiments, p is 1.

In some embodiments, R¹⁴ is selected from hydrogen, fluoro, cyano,hydroxyl, and —OCH₃.

In another aspect, the disclosure features a compound of Formula (III)or a pharmaceutically acceptable salt thereof, wherein:

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and CH; Z′ is selected from

or —CH(R²⁸)—, wherein “1” represents a point of attachment to N(R²⁶);and “2” represents a point of attachment to ring B; ring B is selectedfrom phenyl, pyridinyl, 1H-pyrazolyl, and pyrazinyl; R²¹ is selectedfrom C₃-C₆ cycloalkyl and C₁-C₄ alkyl; R²² is selected from hydrogen andC₁-C₄ alkyl; R²³ is selected from hydrogen and cyano; R²⁴ is selectedfrom hydrogen, hydroxy and halo; R²⁵ is selected from hydrogen, halo,hydroxy, C₁-C₄ alkoxy, —C₁-C₄ alkyl, —C₁-C₄ alkyl-O—C₁-C₄ alkyl, whereineach C₁-C₄ alkyl is optionally substituted with 1 or more substituentsindependently selected from halo and cyano; R²⁶ is selected fromhydrogen and C₁-C₄ alkyl; R²⁷, if present, is independently selectedfrom 1H-pyrazolyl, pyridinyl, and C₁-C₄ alkoxy, wherein the1H-pyrazol-1-yl is optionally substituted with up to 2 substituentsindependently selected from C₁-C₄ alkyl and halo; R²⁸ is selected fromhydrogen and C₁-C₄ alkyl; and o is 0 or 1.

In some embodiments, Z′ is selected from

—CH₂, or —CH(CH₃)—; the portion of the molecule represented by

is selected from

R²¹ is selected from —CH₃ and cyclopropyl; R²² is selected from hydrogenand —CH₃; R²³ is selected from hydrogen and cyano; R²⁴ is selected fromhydrogen, hydroxy and fluoro; R²⁵ is selected from hydrogen, fluoro,hydroxy, —OCH₃, —OCH₂CF₃, —CH₂CH₂OCH₃, —CH₃, —CH₂CH₃, and —CH₂CN; R²⁶ isselected from hydrogen and —CH₃; and R³⁷ is selected from hydrogen,—OCH₃, —OCH₂CH₃, 1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyridin-2-yl. In some embodiments, Z′is —CH₂ or —CH(CH₃)—;

Table 1 below shows the structures of exemplary compounds of thedisclosure.

TABLE 1 Exemplary Compounds # Structure 100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215

216

217

218

219

220

221

222

223

224

225

226

227

228

229

Pharmaceutically acceptable salts of these compounds are alsocontemplated for the uses described herein.

“Pharmaceutically acceptable salt” refers to any salt of a compound ofthe disclosure which retains its biological properties and which is nottoxic or otherwise undesirable for pharmaceutical use. Pharmaceuticallyacceptable salts may be derived from a variety of organic and inorganiccounter-ions well known in the art and include. Such salts include: (1)acid addition salts formed with organic or inorganic acids such ashydrochloric, hydrobromic, sulfuric, nitric, phosphoric, sulfamic,acetic, trifluoroacetic, trichloroacetic, propionic, hexanoic,cyclopentylpropionic, glycolic, glutaric, pyruvic, lactic, malonic,succinic, sorbic, ascorbic, malic, maleic, fumaric, tartaric, citric,benzoic, 3-(4-hydroxybenzoyl)benzoic, picric, cinnamic, mandelic,phthalic, lauric, methanesulfonic, ethanesulfonic,1,2-ethane-disulfonic, 2-hydroxyethanesulfonic, benzenesulfonic,4-chlorobenzenesulfonic, 2-naphthalenesulfonic, 4-toluenesulfonic,camphoric, camphorsulfonic,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic, glucoheptonic,3-phenylpropionic, trimethylacetic, tert-butylacetic, lauryl sulfuric,gluconic, benzoic, glutamic, hydroxynaphthoic, salicylic, stearic,cyclohexylsulfamic, quinic, muconic acid and the like acids; or (2)salts formed when an acidic proton present in the parent compound either(a) is replaced by a metal ion, e.g., an alkali metal ion, an alkalineearth ion or an aluminum ion, or alkali metal or alkaline earth metalhydroxides, such as sodium, potassium, calcium, magnesium, aluminum,lithium, zinc, and barium hydroxide, ammonia or (b) coordinates with anorganic base, such as aliphatic, alicyclic, or aromatic organic amines,such as ammonia, methylamine, dimethylamine, diethylamine, picoline,ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine,arginine, ornithine, choline, N,N′-dibenzylethyl ene-diamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,N-methylglucamine piperazine, tris(hydroxymethyl)-aminomethane,tetramethylammonium hydroxide, and the like. Pharmaceutically acceptablesalts further include, by way of example only, sodium, potassium,calcium, magnesium, ammonium, tetraalkylammonium and the like, and whenthe compound contains a basic functionality, salts of non-toxic organicor inorganic acids, such as hydrochloride, hydrobromide, tartrate,mesylate, besylate, acetate, maleate, oxalate and the like.

Pharmaceutical Compositions

Pharmaceutical compositions of the disclosure comprise one or morecompounds of the disclosure and one or more physiologically orpharmaceutically acceptable carrier. The term “pharmaceuticallyacceptable carrier” refers to a pharmaceutically-acceptable material,composition or vehicle, such as a liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting any subject composition or component thereof. Each carriermust be “acceptable” in the sense of being compatible with the subjectcomposition and its components and not injurious to the patient. Someexamples of materials which may serve as pharmaceutically acceptablecarriers include: (1) sugars, such as lactose, glucose and sucrose; (2)starches, such as corn starch and potato starch; (3) cellulose, and itsderivatives, such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7)talc; (8) excipients, such as cocoa butter and suppository waxes; (9)oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil,olive oil, corn oil and soybean oil; (10) glycols, such as propyleneglycol; (11) polyols, such as glycerin, sorbitol, mannitol andpolyethylene glycol; (12) esters, such as ethyl oleate and ethyllaurate; (13) agar; (14) buffering agents, such as magnesium hydroxideand aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17)isotonic saline; (18) Ringer's solution: (19) ethyl alcohol; (20)phosphate buffer solutions; and (21) other non-toxic compatiblesubstances employed in pharmaceutical formulations.

The compositions of the disclosure may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. In some embodiments, the compositions of the disclosure areadministered orally, intraperitoneally or intravenously. Sterileinjectable forms of the compositions of this disclosure may be aqueousor oleaginous suspension. These suspensions may be formulated accordingto techniques known in the art using suitable dispersing or wettingagents and suspending agents. The sterile injectable preparation mayalso be a sterile injectable solution or suspension in a non-toxicparenterally acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water. Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tween. Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

The pharmaceutically acceptable compositions of this disclosure may beorally administered in any orally acceptable dosage form including, butnot limited to, capsules, tablets, aqueous suspensions or solutions. Inthe case of tablets for oral use, carriers commonly used include lactoseand corn starch. Lubricating agents, such as magnesium stearate, arealso typically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of thisdisclosure may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

The pharmaceutically acceptable compositions of this disclosure may alsobe administered topically, especially when the target of treatmentincludes areas or organs readily accessible by topical application,including diseases of the eye, the skin, or the lower intestinal tract.Suitable topical formulations are readily prepared for each of theseareas or organs. Topical application for the lower intestinal tract canbe effected in a rectal suppository formulation (see above) or in asuitable enema formulation. Topically-transdermal patches may also beused.

For topical applications, the pharmaceutically acceptable compositionsmay be formulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this disclosure include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutically acceptable compositions canbe formulated in a suitable lotion or cream containing the activecomponents suspended or dissolved in one or more pharmaceuticallyacceptable carriers. Suitable carriers include, but are not limited to,mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax,cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The pharmaceutically acceptable compositions of this disclosure may alsobe administered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The amount of the compounds of the present disclosure that may becombined with the carrier materials to produce a composition in a singledosage form will vary depending upon the host treated, the particularmode of administration. Preferably, the compositions should beformulated so that a dosage of between 0.01-100 mg/kg body weight/day ofthe inhibitor can be administered to a patient receiving thesecompositions.

Dosages

Toxicity and therapeutic efficacy of compounds of the disclosure,including pharmaceutically acceptable salts and deuterated variants, canbe determined by standard pharmaceutical procedures in cell cultures orexperimental animals. The LD₅₀ is the dose lethal to 50% of thepopulation. The ED₅₀ is the dose therapeutically effective in 50% of thepopulation. The dose ratio between toxic and therapeutic effects(LD₅₀/ED₅₀) is the therapeutic index. Compounds that exhibit largetherapeutic indexes are preferred. While compounds that exhibit toxicside effects may be used, care should be taken to design a deliverysystem that targets such compounds to the site of affected tissue inorder to minimize potential damage to uninfected cells and, thereby,reduce side effects.

Data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds may lie within a range of circulating concentrations thatinclude the ED₅₀ with little or no toxicity. The dosage may vary withinthis range depending upon the dosage form employed and the route ofadministration utilized. For any compound, the therapeutically effectivedose can be estimated initially from cell culture assays. A dose may beformulated in animal models to achieve a circulating plasmaconcentration range that includes the IC₅₀ (i.e., the concentration ofthe test compound that achieves a half-maximal inhibition of symptoms)as determined in cell culture. Such information can be used to moreaccurately determine useful doses in humans. Levels in plasma may bemeasured, for example, by high performance liquid chromatography.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of a compound of the present disclosure in the composition willalso depend upon the particular compound in the composition.

Treatment

In an aspect, the disclosure features a method for inhibiting RETactivity in a cell or in a patient, comprising the step of contactingthe cell or administering to the patient a compound as described hereinor a pharmaceutical composition as described herein.

In another aspect, the disclosure features a method for treating asubject suffering from a condition mediated by aberrant RET activity,comprising administering to the subject a therapeutically effectiveamount of a compound as described herein or a pharmaceutical compositionas described herein.

In another aspect, the disclosure features a method for treating asubject who has developed resistance to a treatment for a conditionmediated by aberrant RET activity, comprising administering to thesubject a therapeutically effective amount of a compound as describedherein or a pharmaceutical composition as described herein.

RET fusions have been implicated in several types of cancers. Generally,these RET fusions have a RET kinase domain that is the same as inwild-type RET; therefore, as used herein, any RET protein with the samekinase domain as wild-type RET will be referred to as “wild-type RET”unless noted otherwise. Mutations can occur at least in the RETextracellular and kinase domains. Mutations can occur in the RET kinasedomain, leading to resistant mutants of RET.

The activity of exemplary compounds that are approved or in developmentfor RET-related conditions is shown below. As shown, the compounds areactive against the wild-type RET, but are much less active against themutated forms (“wild-type RET inhibitors”).

TABLE 2 RET wt RET V804L RET V804M Biochemical Biochemical BiochemicalCompound IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM) Cabozantinib 46 219 585Vandetanib 1.2 902 607 Sorafenib 7.9 95.2 32.4 Regorafenib 5.1 29.8 46.7

The disclosure provides compounds that inhibit both wild-type RET andresistant mutants of RET. In addition, the compounds of the disclosurecan be selective for wild-type RET, over other kinases, thus leading toreduced toxicities associated with inhibiting other kinases. In oneaspect, compounds of the disclosure are selective for RET over KDR. Inone aspect, compounds of the disclosure do not cause adverse effectssuch as hypertension, arterial thrombosis, and hemorrhage.

In addition, the disclosure provides inhibitors of mutant RET. Mutationscan be predicted using structural biology and computational analyses, aswell as by examining codon sequences in which a sequence change givesrise to a codon for a different amino acid. Using such methods,resistant mutants for RET are predicted to have point mutations at the804 gatekeeper residue in the RET protein and/or at residues at or nearthe gatekeeper residue. In some embodiments, the mutation may be at oneor more of the 804, 806, 810, 865, 870, 891, and 918 residues. Specificexamples of RET resistant mutants include: V804L, V804M, V804E, Y806C,Y806S, Y806H, Y806N, G810R, G810S, L865V, L870F, S891A and M918Tmutants.

Mutations occurring from administration of a particular inhibitor (e.g.,a known RET wild-type inhibitor) can be determined experimentally byexposing cells to a mutation-promoting agent, such as ENU. The cells arewashed, then plated with increasing concentrations (2-100× proliferationIC₅₀) of the compound of choice. The wells with cellular outgrowth arethen collected after 3-4 weeks. The RET kinase domain is then sequencedto identify resistance mutations (i.e., altered forms of the RET proteinthat retain enzymatic activity). Resistance can be confirmed by exposingthese cells with the compound of choice. Resistant mutants that havebeen identified experimentally include the V804L, V804E, V804M, andY806H mutants. In some embodiments, the mutation is a substitution ofcysteine (C609, C611, C618, C620, C630, and C634) in the RETextracellular domain for any other amino acid. In some embodiments, theRET cysteine variants (affecting C609, C611, C618, and C620) are the“Janus mutations.” In some embodiments, RET mutations include: RETC634W, RET M918T, V804L, V804E, V804M, V804L, Y806C, Y806S, Y806N,Y806H, G810R, G810S, L865V, L870F, and S891A mutants.

Because of their activity against wild-type RET and mutant RET, thecompounds described herein can be used to treat a patient with acondition associated with aberrant RET activity. The compounds describedherein can provide treatments for irritable bowel syndrome (IBS),proliferative diseases, and any other conditions related to aberrant RETactivity. The compounds can be used to treat irritable bowel syndrome.The compounds can be used to treat various cancers. In some embodiments,the cancer is selected from papillary thyroid carcinoma (PTC), medullarythyroid cancer (MTC), pheochromocytoma (PC), pancreatic ductaladenocarcinoma, multiple endocrine neoplasia (MEN2A and MEN2B),metastatic breast cancer, testicular cancer, small cell lung cancer,non-small cell lung cancer, chronic myelomonocytic leukemia, colorectalcancer, ovarian cancer, and cancers of the salivary gland. In someembodiments, the cancer is a solid tumor. In some embodiments, thecondition associated with aberrant RET activity is a thyroid cancer(e.g., papillary thyroid carcinoma, thyroid adenocarcinoma, or MTC,e.g., familial MTC), lung cancer (e.g., lung adenocarcinoma, small-celllung carcinoma, or non-small cell lung carcinoma), breast cancer (e.g.,estrogen receptor-positive tumors and endocrine-resistant tumors e.g.,resistant to oestrogen modulators such as tamoxifen, agents that blockoestrogen biosynthesis such as aromatase inhibitors, and oestrogenreceptor antagonists such as fulvestrant), pancreatic cancer (e.g.,carcinoma of the pancreas or pancreatic ductal carcinoma),haematopoietic cancer, e.g., a leukemia (e.g., chronic myelomonocyticleukemia or acute myeloid leukemia), colon cancer (e.g., coloncarcinoma), melanoma (e.g., cutaneous or desmoplastic malignantmelanomas), prostate cancer, renal cancer (e.g., renal cell carcinoma),and head and neck tumors, neuroblastoma, ganglioneuroma (e.g.,ganglioneuroma of the mouth or gut), colon cancer (e.g., sporadic coloncancers), MEN2A (multiple endocrine neoplasia type 2A), or MEN2B(multiple endocrine neoplasia type 2B). In one aspect, the MEN2A ischaracterized by MTC and includes adrenal tumor pheochromocytoma.Substitutions of cysteines in RET are found in subjects with MEN2A andalso frequent in FMTC. RET extracellular domain exon 8 mutations, suchas G533C) or the RET intracellular domain (residues E768, L790, Y791,V804, and S891) are associated with FMTC or MEN2A. Substitutions in theRET kinase domain, Met918 to Thr (M918T) or A883F are found in subjectswith MEN2B. RET M918T and RET A883F are also found in sporadic MTC.

The compounds can also be used to treat a patient who has developedresistance to a wild-type RET inhibitor, or a patient with a particularRET mutant. The method includes the step of administering a compound orcomposition of the disclosure that is active against one or more RETresistant mutants. In certain embodiments, the RET resistant mutant isselected from V804L, V804M, V804E, Y806C, Y806S, Y806N, Y806H, G810R,G810S, L865V, L870F, S891A and M918T. By “active” is meant that acompound has an IC₅₀ of less than 1 μM, 500 nM, 250 nM, 100 nM, 75 nM,50 nM, 25 nM, 10 nM, or 5 nM when measured in a biochemical assay,against at least one resistant mutant.

In some embodiments, the cancer is a solid tumor. In some embodiments,the condition associated with aberrant RET activity is a thyroid cancer(e.g., papillary thyroid carcinoma, thyroid adenocarcinoma, or MTC,e.g., familial MTC), lung cancer (e.g., lung adenocarcinoma, small-celllung carcinoma, or non-small cell lung carcinoma), breast cancer (e.g.,estrogen receptor-positive tumors and endocrine-resistant tumors e.g.,resistant to oestrogen modulators such as tamoxifen, agents that blockoestrogen biosynthesis such as aromatase inhibitors, and oestrogenreceptor antagonists such as fulvestrant), pancreatic cancer (e.g.,carcinoma of the pancreas or pancreatic ductal carcinoma),haematopoietic cancer, e.g., a leukemia (e.g., chronic myelomonocyticleukemia or acute myeloid leukemia), colon cancer (e.g., coloncarcinoma), melanoma (e.g., cutaneous or desmoplastic malignantmelanomas), prostate cancer, renal cancer (e.g., renal cell carcinoma),and head and neck tumors, neuroblastoma, ganglioneuroma (e.g.,ganglioneuroma of the mouth or gut), colon cancer (e.g., sporadic coloncancers), MEN2A (multiple endocrine neoplasia type 2A), or MEN2B(multiple endocrine neoplasia type 2B). In one aspect, the MEN2A ischaracterized by MTC and includes adrenal tumor pheochromocytoma.Substitutions of cysteines in RET are found in subjects with MEN2A andalso frequent in FMTC. RET extracellular domain exon 8 mutations, suchas G533C) or the RET intracellular domain (residues E768, L790, Y791,V804, and S891) are associated with FMTC or MEN2A. Substitutions in theRET kinase domain, Met918 to Thr (M918T) or A883F are found in subjectswith MEN2B. RET M918T and RET A883F are also found in sporadic MTC.

The compounds may also be used to treat a subject having a RET-alteredcell, cancer, gene, or gene product. The RET alteration may be, e.g., apoint mutation, insertion, deletion, amplification, or fusion, or acombination thereof.

The compounds may also be used to treat a subject having a RET-alteredcell, cancer, gene, or gene product comprising a RET alterationdescribed in Table 3 or Table 4 herein. In some embodiments, the subjecthas a fusion between RET and a RET fusion partner listed in Table 3,e.g., comprises a fusion protein that comprises RET or a fragmentthereof and a protein of Table 3 or fragment thereof. In someembodiments, the fusion partner is N-terminal or C-terminal of RET. Insome embodiments, the subject has an alteration at a position in RETthat is described in Table 4. In some embodiments, a subset of thesubject's cells, e.g., a subset of the subject's tumor cells, comprisethe RET alteration. In some embodiments, a subset of the subject'scells, e.g., a subset of the subject's tumor cells, are RET-altered. Insome embodiments, the subject has a cancer listed in Table 3, e.g., thesubject has both a RET mutation and a cancer listed in Table 3.

TABLE 3 RET fusions RET fusion partner Exemplary cancers in which thefusion is found BCR Chronic Myelomonocytic Leukemia (CMML) CLIP 1Adenocarcinoma KIFSB NSCLC, Ovarian Cancer, Spitzoid Neoplasm; LungAdenocarcinoma, Adenosquamous Carcinomas CCDC6 NSCLC, Colon Cancer,Papillary Thyroid Cancer; Adenocarcinoma; Lung Adenocarcinoma;Metastatic Colorectal Cancer; Adenosquamous Carcinoma, Metastaticpapillary thyroid cancer PTClex9 Metastatic papillary thyroid cancerNCOA4 Papillary Thyroid Cancer, NSCLC, Colon Cancer, Salivary GlandCancer, Metastatic Colorectal Cancer; Lung Adenocarcinoma, AdenosquamousCarcinomas; Diffuse Sclerosing Variant of Papillary Thyroid CancerTRIM33 NSCLC, Papillary Thyroid Cancer ERC1 Papillary Thyroid Cancer,Breast Cancer FGFRIOP CMML, Primary Myelofibrosis with secondary AcuteMyeloid Leukemia MBD1 Papillary Thyroid Cancer RAB61P2 Papillary ThyroidCancer PRKAR1A Papillary Thyroid Cancer TRIM24 Papillary Thyroid CancerKTN1 Papillary Thyroid Cancer GOLGA5 Papillary Thyroid Cancer, SpitzoidNeoplasms HOOK3 Papillary Thyroid Cancer KIAA1468 Papillary ThyroidCancer, Lung Adenocarcinoma TRIM27 Papillary Thyroid Cancer AKAP13Papillary Thyroid Cancer FKBP15 Papillary Thyroid Cancer SPECC1LPapillary Thyroid Cancer, Thyroid Gland Carcinoma TBL1XR1 PapillaryThyroid Cancer, Thyroid Gland Carcinoma CEP55 Diffuse Gastric CancerCUX1 Lung Adenocarcinoma ACBD5 Papillary Thyroid Carcinoma MYH13Medullary Thyroid Carcinoma PIBF1 Bronchiolus Lung Cell CarcinomaKIAA1217 Papillary Thyroid Cancer, Lung Adenocarcinoma, NSCLC MPRIPNSCLC

TABLE 4 RET mutations Amino acid position 2 Amino acid position 3 Aminoacid position 4 Amino acid position 5 Amino acid position 6 Amino acidposition 7 Amino acid position 8 Amino acid position 11 Amino acidposition 12 Amino acid position 13 Amino acid position 20 Amino acidposition 32 (e.g., S32L) Amino acid position 34 (e.g., D34S) Amino acidposition 40 (e.g., L40P) Amino acid position 64 (e.g., P64L) Amino acidposition 67 (e.g., R67H) Amino acid position 114 (e.g., R114H) Aminoacid position 136 (e.g., glutamic acid to stop codon) Amino acidposition 145 (e.g., V145G) Amino acid position 180 (e.g., arginine tostop codon) Amino acid position 200 Amino acid position 292 (e.g.,V292M) Amino acid position 294 Amino acid position 321 (e.g., G321R)Amino acid position 330 (e.g., R330Q) Amino acid position 338 (e.g.,T338I) Amino acid position 360 (e.g., R360W) Amino acid position 373(e.g., alanine to frameshift) Amino acid position 393 (e.g., F393L)Amino acid position 432 Δ Amino acid residues 505-506 (6-Base PairIn-Frame Germline Deletion in Exon 7) Amino acid position 510 (e.g.,A510V) Amino acid position 511 (e.g., E511K) Amino acid position 513(e.g., A513D) Amino acid position 515 (e.g., C515S, C515W) Amino acidposition 525 (e.g., R525W) Amino acid position 531 (e.g., C531R, or 9base pair duplication) Amino acid position 532 (e.g., duplication) Aminoacid position 533 (e.g., G533C or G533S) Amino acid position 550 (e.g.,G550E) Amino acid position 591 (e.g., V591I) Amino acid position 593(e.g., G593E) Amino acid position 600 (e.g., R600Q) Amino acid position602 (e.g., I602V) Amino acid position 603 (e.g., K603Q or K603E2) Aminoacid position 606 (e.g., Y606C) Amino acid position 609 (e.g., C609Y,C609S, C609G, C609R, C609F, or C609W) Amino acid position 611 (e.g.,C611R, C611S, C611G, C611Y, C611F, or C611W) Amino acid position 618(e.g., C618S, C618Y, C618R, C618Y, C618G, C618F, C618W) Amino acidposition 619 (e.g., F619F) Amino acid position 620 (e.g., C620S, C620W,C620R, C620G, C620L, C620Y, C620F) Amino acid position 623 (e.g., E623K)Amino acid position 624 (e.g., D624N) Amino acid position 630 (e.g.,C630A, C630R, C630S, C630Y, or C630F) Amino acid position 631 (e.g.,D631N, D631Y, D631A, D631G, D631V, or D631E) Amino acid position 632(e.g., E632K or E632G5) Δ Amino acid residues 632-633 (6-Base PairIn-Frame Germline Deletion in Exon 11) Amino acid position 633 (e.g., 9base pair duplication) Amino acid position 634 (e.g., C634W, C634Y,C634S, C634R, C634F, C634G, C634L, C634A, or C634T, or an insertionELCR2, or a 12 base pair duplication) Amino acid position 635 (e.g.,R635G) Amino acid position 636 (e.g., T636P or T636M4) Amino acidposition 640 (e.g., A640G) Amino acid position 641 (e.g., A641S orA641T8) Amino acid position 648 (e.g., V6481) Amino acid position 649(e.g., S649L) Amino acid position 664 (e.g., A664D) Amino acid position665 (e.g., H665Q) Amino acid position 666 (e.g., K666E, K666M, or K666N)Amino acid position 686 (e.g., S686N) Amino acid position 691 (e.g.,G691S) Amino acid position 694 (e.g., R694Q) Amino acid position 700(e.g., M700L) Amino acid position 706 (e.g., V706M or V706A) Amino acidposition 713 splice variant (e.g., E713K) Amino acid position 736 (e.g.,G736R) Amino acid position 748 (e.g., G748C) Amino acid position 750(e.g., A750P) Amino acid position 765 (e.g., S765P) Amino acid position766 (e.g., P766S or P766M6) Amino acid position 768 (e.g., E768Q orE768D) Amino acid position 769 (e.g., L769L) Amino acid position 770(e.g., R770Q) Amino acid position 771 (e.g., D771N) Amino acid position777 (e.g., N777S) Amino acid position 778 (e.g., V778I) Amino acidposition 781 (e.g., Q781R) Amino acid position 790 (e.g., L790F) Aminoacid position 791 (e.g., Y791F or Y791N) Amino acid position 802 Aminoacid position 804 (e.g., V804L, V804M, V804M*, or V804E) Amino acidposition 805 (e.g., E805K) Amino acid position 806 (e.g., E806C, Y806E,Y806F, Y806S, Y806G, or Y806C) Amino acid position 818 (e.g., E818K)Amino acid position 819 (e.g., S819I) Amino acid position 823 (e.g.,G823E) Amino acid position 826 (e.g., Y826M) Amino acid position 833(e.g., R833C) Amino acid position 841 (e.g., P841L or P841P) Amino acidposition 843 (e.g., E843D) Amino acid position 844 (e.g., R844W, R844Q,or R844L) Amino acid position 848 (e.g., M848T) Amino acid position 852(e.g., 1852M) Amino acid position 866 (e.g., A866W) Amino acid position873 (e.g., R873W) Amino acid position 876 (e.g., A876V) Amino acidposition 881 (e.g., L881V) Amino acid position 882 Amino acid position883 (e.g., A883F, A883S, A883T, or A883T*) Amino acid position 884(e.g., E884K) Amino acid position 886 (e.g., R886W) Amino acid position891 (e.g., S891A) Amino acid position 897 (e.g., R897Q) Amino acidposition 898 (e.g., D898V) Amino acid position 901 (e.g., E901K) Aminoacid position 904 (e.g., S904F or S904C2) Amino acid position 907 (e.g.,K907E or K907M) Amino acid position 908 (e.g., R908K) Amino acidposition 911 (e.g., G911D) Amino acid position 912 (e.g., R912P, R912Q)Amino acid position 918 (e.g., M918T, M918V, or M918L6) Amino acidposition 919 (e.g., A919V) Amino acid position 921 (e.g., E921K) Aminoacid position 922 (e.g., S922P or S922Y) Amino acid position 930 (e.g.,T930M) Amino acid position 961 (e.g., F961L) Amino acid position 972(e.g., R972G) Amino acid position 982 (e.g., R982C) Amino acid position1009 (e.g., M1009V) Amino acid position 1017 (e.g., D1017N) Amino acidposition 1041 (e.g., V1041G) Amino acid position 1064 (e.g., M1064T)RET + 3

RET has two primary protein and mRNA isoforms, named RET51 and RET9. Insome embodiments, RET has a sequence of isoform RET51 (SEQ ID NO: 1).The kinase domain corresponds to amino acids 724-1016 of SEQ ID NO: 1.

In some embodiments, RET has a sequence of isoform RET9 (SEQ ID NO: 2).

In some embodiments, RET51 is encoded by a nucleic acid having thesequence of SEQ ID NO: 3.

In some embodiments, RET9 is encoded by a nucleic acid having thesequence of SEQ ID NO: 4.

The compounds and compositions described herein can be administeredalone or in combination with other compounds, including otherRET-modulating compounds, or other therapeutic agents. In someembodiments, the compound or composition of the disclosure may beadministered in combination with one or more compounds selected fromcabozantinib (COMETRIQ), vandetanib (CALPRESA), sorafenib (NEXAVAR),sunitinib (SUTENT), regorafenib (STAVARGA), ponatinib (ICLUSIG),bevacizumab (AVASTIN), crizotinib (XALKORI), or gefitinib (IRESSA). Thecompound or composition of the disclosure may be administeredsimultaneously or sequentially with the other therapeutic agent by thesame of different routes of administration. The compound of thedisclosure may be included in a single formulation with the othertherapeutic agent or in separate formulations.

Synthesis

Compounds of the disclosure, including salts and N-oxides thereof, canbe prepared using known organic synthesis techniques and can besynthesized according to any of numerous possible synthetic routes, suchas those in the Schemes below. The reactions for preparing compounds ofthe disclosure can be carried out in suitable solvents which can bereadily selected by one of skill in the art of organic synthesis.Suitable solvents can be substantially non-reactive with the startingmaterials (reactants), the intermediates, or products at thetemperatures at which the reactions are carried out, e.g., temperatureswhich can range from the solvent's freezing temperature to the solvent'sboiling temperature. A given reaction can be carried out in one solventor a mixture of more than one solvent. Depending on the particularreaction step, suitable solvents for a particular reaction step can beselected by the skilled artisan.

Preparation of compounds of the disclosure can involve the protectionand deprotection of various chemical groups. The need for protection anddeprotection, and the selection of appropriate protecting groups, can bereadily determined by one skilled in the art. The chemistry ofprotecting groups can be found, for example, in Wuts and Greene,Protective Groups in Organic Synthesis, 4th ed., John Wiley & Sons: NewJersey, (2006), which is incorporated herein by reference in itsentirety.

Reactions can be monitored according to any suitable method known in theart. For example, product formation can be monitored by spectroscopicmeans, such as nuclear magnetic resonance (NMR) spectroscopy (e.g., ¹Hor ¹³C), infrared (IR) spectroscopy, spectrophotometry (e.g.,UV-visible), mass spectrometry (MS), or by chromatographic methods suchas high performance liquid chromatography (HPLC) or thin layerchromatography (TLC). Analytical instruments and methods for compoundcharacterization:

LC-MS:

Unless otherwise indicated, all liquid chromatography-mass spectrometry(LC-MS) data (sample analyzed for purity and identity) were obtainedwith an Agilent model-1260 LC system using an Agilent model 6120 massspectrometer utilizing ES-API ionization fitted with an Agilent Poroshel120 (EC-C18, 2.7 um particle size, 3.0×50 mm dimensions) reverse-phasecolumn at 22.4 degrees Celsius. The mobile phase consisted of a mixtureof solvent 0.1% formic acid in water and 0.1% formic acid inacetonitrile. A constant gradient from 95% aqueous/5% organic to 5%aqueous/95% organic mobile phase over the course of 4 minutes wasutilized. The flow rate was constant at 1 mL/min.

Prep LC-MS:

Preparative HPLC was performed on a Shimadzu Discovery VP® Preparativesystem fitted with a Luna 5u C18(2) 100A, AXIA packed, 250×21.2 mmreverse-phase column at 22.4 degrees Celsius. The mobile phase consistedof a mixture of solvent 0.1% formic acid in water and 0.1% formic acidin acetonitrile. A constant gradient from 95% aqueous/5% organic to 5%aqueous/95% organic mobile phase over the course of 25 minutes wasutilized. The flow rate was constant at 20 mL/min. Reactions carried outin a microwave were done so in a Biotage Initiator microwave unit.

Silica Gel Chromatography:

Silica gel chromatography was performed on either a Teledyne IscoCombiFlash® Rf unit or a Biotage® Isolera Four unit.

Proton NMR:

Unless otherwise indicated, all ¹H NMR spectra were obtained with aVarian 400 MHz Unity Inova 400 MHz NMR instrument (acquisition time=3.5seconds with a 1 second delay; 16 to 64 scans). Where characterized, allprotons were reported in DMSO-d6 solvent as parts-per million (ppm) withrespect to residual DMSO (2.50 ppm).

EXAMPLES

The following examples are intended to be illustrative, and are notmeant in any way to be limiting.

The below Synthetic Protocols and specific synthesis examples are meantto provide general guidance in connection with preparing the compoundsof the disclosure. One skilled in the art would understand that thepreparations shown in the Synthetic Protocols and specific examples canbe modified or optimized using general knowledge of organic chemistry toprepare various compounds of the disclosure.

In general, the compounds of this disclosure were prepared using one ofthe following five Synthetic Protocols.

Synthetic Protocol 1:

A heteroaryl dihalide can be coupled to an amino pyrazole undernucleophilic aromatic substitution reaction conditions using a base suchas diisopropylethylamine (DIPEA) or triethylamine (TEA) in a polarsolvent to provide the bicyclic ring system. The bicyclic heteroarylhalide can then undergo an SnAr reaction with an amino ester or aminoacid intermediate to give a tricyclic ring system. The amino ester oramino acid intermediates are either commercially available or preparedas described in “Synthesis of Heterocycle Intermediates”. When an aminoacid is used, the tricyclic acid product can then be coupled to avariety amines under amide coupling reaction conditions to give thefinal product examples. The amines are either commercially available, orcould be prepared as described below under the heading “Synthesis ofAmine Intermediates”. Alternatively, if an amino ester was used in theSnAr reaction, the tricyclic ester is hydrolyzed under basic, acidic, orother conditions to give the tricyclic acid intermediate. The tricyclicacid intermediate can be coupled to an amine as described above to thengive the final compound.

Synthetic Protocol 2:

A heteroaryl dihalide can be coupled to an amino ester (eithercommercially available or prepared as described in “Synthesis ofHeterocycle Intermediates”) under SnAr conditions, using a base such asDIPEA, TEA, Cs₂CO₃, CsF, Na₂CO₃, or other bases. The resulting bicyclicheteroaryl halide can then be coupled with an amino pyrazole undernucleophilic aromatic substitution reaction conditions or palladiummediated coupling conditions. The tricyclic ester can then be hydrolyzedunder basic, acidic, or other conditions to give a tricyclic acidintermediate. The final compound examples are then prepared by an amidecoupling reaction with the tricyclic acid and an amine. The amines couldbe commercially available, or could be prepared as described below underthe heading “Synthesis of Amine Intermediates”.

Synthetic Protocol 3:

An N-protected amino acid intermediate can be coupled to an amine usingamide coupling reaction conditions. The protected amino acidintermediate starting material used above could be commerciallyavailable, or prepared as described in “Synthesis of HeterocycleIntermediates” below. The protecting group can be benzyl,tert-butoxycarbonyl, or others. The amines used in the amide couplingare either commercially available, or could be prepared as describedbelow under the heading “Synthesis of Amine Intermediates”. Theprotecting group is then removed under typical deprotection conditionsto give an amino amide intermediate. The amino amide intermediate isthen coupled to a bicyclic heteroaryl halide under SnAr conditions,using a base such as DIPEA, TEA, Cs₂CO₃, CsF, Na₂CO₃, or other bases, togive the final compound. In some instances, a palladium mediatedcoupling reaction could be used instead of a SnAr reaction.

Synthetic Protocol 4:

An N-protected amino acid intermediate can be coupled to an amine usingamide coupling reaction conditions. The protected amino acidintermediate starting material used above could be commerciallyavailable, or prepared as described in “Synthesis of HeterocycleIntermediates” below. The protecting group can be benzyl,tert-butoxycarbonyl, or others. The amines used in the amide couplingare either commercially available, or could be prepared as describedbelow under the heading “Synthesis of Amine Intermediates”. Theprotecting group is then removed under typical deprotection conditionsto give an amino amide intermediate. The amino amide intermediate isthen coupled to a heteroaryl dihalide under SnAr conditions, using abase such as DIPEA, TEA, Cs₂CO₃, CsF, Na₂CO₃, or other bases, to give aheteroaryl halide. The heteroaryl halide is then coupled to an aminopyrazole intermediate under SnAr or palladium mediated couplingconditions to give the final compound.

Synthetic Protocol 5:

An amino acid intermediate can be coupled to a heteroaryl dihalide underSnAr conditions using DIPEA, TEA, Cs₂CO₃, CsF, Na₂CO₃, or other bases.The resulting acid intermediate can then be coupled to an amine usingamide coupling reaction conditions. The amines used in the amidecoupling are either commercially available, or could be prepared asdescribed below under the heading “Synthesis of Amine Intermediates”.The amide product, which contains a heteroaryl halide group, is thencoupled to an aminopyrazole intermediate under palladium mediatedcoupling conditions to give the final compound.

Example 1. Synthesis of Compound 161 Step 1: Synthesis of2-chloro-6-methyl-N-(5-methyl-1H-pyrazol-3-yl)pyrimidin-4-amine

A suspension of 2,4-dichloro-6-methyl-pyrimidine (120.00 g 736.2 mmol,1.00 eq), 5-methyl-1H-pyrazol-3-amine (78.65 g, 0.81 mol, 1.10 eq) andDIPEA (142.72 g, 1.10 mol, 1.50 eq) in DMSO (400 mL) was heated at 60°C. for 16 hrs. TLC (PE/EA, 5:1, 1:1) showed the reaction was complete.The reaction mixture was cooled to 30° C. and poured into ice-water (800mL). The resulting mixture was extracted with MTBE (800 mL×10). Thecombined organic layers were washed with water (400 mL×3), brine (400mL×3) and dried over Na₂SO₄. After filtration, the filtrate wasconcentrated under reduced pressure and the residue was recrystallizedfrom DCM (10 mL/g) to afford2-chloro-6-methyl-N-(5-methyl-1H-pyrazol-3-yl)pyrimidin-4-amine (105.60g, 472.14 mmol, 64%) as a yellow solid. The structure was confirmed byLC-MS and NMR.

Step 2: Synthesis of4-methoxy-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylicacid

A mixture of 4-methoxypiperidine-4-carboxylic acid (214 mg, 1.34 mmol),2-chloro-6-methyl-N-(5-methyl-1H-pyrazol-3-yl)pyrimidin-4-amine (300 mg,1.34 mmol) and DIPEA (520 mg, 4.02 mmol) in 2-propanol (3 mL) was heatedto 150° C. in a microwave reactor. The reaction mixture was stirred for50 min, and then was cooled to ambient temperature. The reaction mixturewas concentrated and the residue was purified by column chromatographyon silica gel (gradient elution, 0 to 10% methanol-DCM) to give4-methoxy-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylicacid (435 mg, 94%). MS (ES+) C₁₆H₂₂N6O₃ requires: 346, found: 347[M+H]⁺.

Step 3: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxy-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)piperidine-4-carboxamide

HBTU (220 mg, 0.577 mmol) was added to a mixture of4-methoxy-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylicacid (100 mg, 0.289 mmol), (6-(1H-pyrazol-1-yl)pyridin-3-yl)methanamine(50 mg, 0.289 mmol) and DIPEA (112 mg, 0.866 mmol) in DMF (3.0 mL) atambient temperature. The reaction mixture was stirred for 16 h, and thenwas partitioned between dichloromethane and water. The dichloromethanelayer was washed with saturated aqueous sodium chloride solution, driedover sodium sulfate, filtered, and concentrated. The residue waspurified by column chromatography on silica gel (gradient elution, 0 to10%0/methanol-DCM) to give the title compound (35 mg, 24%) as a whitesolid.

Example 2. Synthesis of Compound 166 Step 1: Synthesis of methyl3-ethyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxylate

A solution of methyl 3-ethylazetidine-3-carboxylate (50 mg, 0.40 mmol),2-chloro-6-methyl-N-(5-methyl-1H-pyrazol-3-yl)pyrimidin-4-amine (78 mg,0.40 mmol) and DIPEA (150 mg, 1.20 mmol) in isoamyl alcohol (0.5 mL) wasstirred at 150° C. for 5 h. The reaction mixture was then cooled toambient temperature and concentrate to give a residue (85 mg) which wasused in the next step without further purification.

Step 2: Synthesis of3-ethyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxylicacid

LiOH (380 mg, 9.00 mmol) was added to a solution of methyl3-ethyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxylate(600 mg, 1.80 mmol) in methanol (4 mL), THF (4 mL) and water (4 mL) atambient temperature. The reaction mixture was stirred for 4 h, and thenwas concentrated. The residue was dissolved in DMSO and purified byreverse-phase HPLC (ACN/H2O gradient elution with 0.1% TFA) to give3-ethyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxylicacid (550 mg).

Step 3: Synthesis of(S)-3-ethyl-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxamide

A mixture of3-ethyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)azetidine-3-carboxylicacid (67 mg, 0.21 mmol),(S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethanamine hydrochloride(51 mg, 0.21 mmol), HBTU (80 mg, 0.21 mmol) and NMM (64 mg, 0.63 mmol)in DMF (2 mL) was stirred at 25° C. for 1 h. The solution was purifiedby preparative HPLC to give the title product (71 mg, 66%) as a whitesolid.

Example 3. Synthesis of Compound 107 Step 1: Synthesis of methyl1-(6-bromo-4-methylpyridin-2-yl)-3-methylazetidine-3-carboxylate

A mixture of 2-bromo-6-fluoro-4-methylpyridine (500 mg, 2.63 mmol),methyl 3-methylazetidine-3-carboxylate (480 mg, 2.63 mmol), Na₂CO3 (840mg, 7.89 mmol), and isoamyl alcohol (0.5 mL) was heated to 140° C. for 8h. The reaction mixture was then cooled to ambient temperature, andmethanol (15 mL) was added. The mixture was filtered, and the filtratewas concentrated to give methyl1-(6-bromo-4-methylpyridin-2-yl)-3-methylazetidine-3-carboxylate (700mg) which was used in the next step without further purification.

Step 2: Synthesis of methyl3-methyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)azetidine-3-carboxylate

A mixture of methyl1-(6-bromo-4-methylpyridin-2-yl)-3-methylazetidine-3-carboxylate (400mg, 1.27 mmol), 5-methyl-1H-pyrazol-3-amine (119 mg, 1.52 mmol),Pd₂(dba)₃ (116 mg, 0.127 mmol), Xantphos (147 mg, 0.254 mmol) and KOAc(373 mg, 3.810 mmol) in 1,4-dioxane (15 mL) under nitrogen was stirredat 100° C. for 8 h. The mixture was cooled to ambient temperature andpartitioned between ethyl acetate and water. The organic layer waswashed with saturated aqueous sodium chloride solution, and the washedsolution was dried over sodium sulfate. The dried solution was filtered,and the filtrate was concentrated to afford methyl3-methyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)azetidine-3-carboxylate(700 mg), which was used in the next step without further purification.

Steps 3-4: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-3-methyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)azetidine-3-carboxamide

The title compound was prepared from of methyl3-methyl-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)azetidine-3-carboxylateusing the same two-step procedure (hydrolysis and amide coupling) as inSynthetic Protocol 1 to give a white solid.

Example 4. Synthesis of Compound 136 Step 1: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-benzyl-4-hydroxypiperidine-4-carboxamide

A mixture of 1-benzyl-4-hydroxypiperidine-4-carboxylic acid (436 mg,1.85 mmol), (6-(1H-pyrazol-1-yl)pyridin-3-yl)methanamine (0.323 g, 1.85mmol), HATU (1.05 g, 2.78 mmol) and DIPEA (717 mg, 5.56 mmol) in DMF (20mL) was stirred at ambient temperature for 16 h. The reaction mixturewas then partitioned between ethyl acetate and water. The layers wereseparated, and the aqueous layer was further extracted with ethylacetate. The organic layers were combined, and the combined layers werewashed sequentially with water, brine, and then dried over sodiumsulfate. The dried solution was filtered, and the filtrate wasconcentrated. The residue was purified by flash column chromatography onsilica gel (gradient elution, 0 to 10% methanol-DCM to giveN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-benzyl-4-hydroxypiperidine-4-carboxamide(218 mg, yield 30%) as a yellow solid. MS (ES+) C₂₂H₂₅N₅O₂ requires:391, found 392 [M+H]⁺.

Step 2: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-hydroxypiperidine-4-carboxamide

To a solution ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-benzyl-4-hydroxypiperidine-4-carboxamide(200 mg, 0.510 mmol) in MeOH (5 mL) was added Pd/C (1-% by weight, 100mg) under N₂. The resulting suspension was evacuated and refilled withhydrogen three times and stirred at ambient temperature for 16 h underhydrogen. The reaction mixture was then filtered through celite, and thefiltrate was concentrated to giveN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-hydroxypiperidine-4-carboxamide(150 mg), which was used in the next step without purification. MS (ES+)C₁₆H₂₀N₄O₂ requires: 300, found: 301 [M+H]⁺.

Step 3: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-hydroxy-1-(4-methyl-6-(3-methyl-1H-pyrazol-5-ylamino)pyrimidin-2-yl)piperidine-4-carboxamide

A mixture ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-hydroxypiperidine-4-carboxamide(100 mg, 0.33 mmol),2-chloro-6-methyl-N-(3-methyl-1H-pyrazol-5-yl)pyrimidin-4-amine (70 mg,0.33 mmol) and K₂CO₃(96 mg, 0.70 mmol) in DMF (10 mL) was heated to 80°C. for 16 h. The mixture was slowly poured into ice water and extractedwith ethyl acetate (3×). The combined organic layers were washedsequentially with water, brine, and then dried over sodium sulfate. Thedried solution was filtered, and the filtrate was concentrated. Theresidue was purified by flash column chromatography on silica gel(gradient elution, 0 to 10% methanol-DCM to afford the title compound(100 mg, yield 62%) as a yellow solid.

Example 5. Synthesis of Compound 160 Step 1: Synthesis of tert-butyl4-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methylcarbamoyl)-4-methoxypiperidine-1-carboxylate

To a solution of1-(tert-butoxycarbonyl)-4-methoxypiperidine-4-carboxylic acid (200 mg,0.771 mmol), (6-(1H-pyrazol-1-yl)pyridin-3-yl)methanamine (148 mg, 0.848mmol) and DIPEA (298 mg, 2.31 mmol) in THF (10 mL) was added HATU (293mg, 0.770 mmol) at ambient temperature. The solution was stirred atambient temperature for 18 h, and then was concentrated. The residue waspurified by flash column chromatography on silica gel (gradient elution,0 to 10% methanol-DCM) to give tert-butyl4-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methylcarbamoyl)-4-methoxypiperidine-1-carboxylate(300 mg, yield 94%) as a yellow oil. MS (ES+) C₂₁H₂₉N₅O₄ requires: 415found: 416 [M+H]⁺.

Step 2: Synthesis ofN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxypiperidine-4-carboxamidehydrochloride

To a solution of tert-butyl4-(((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamoyl)-4-methoxypiperidine-1-carboxylate(340 mg, 0.818 mmol) in dioxane (5 mL) was added HCl (4 M in dioxane, 5mL) at ambient temperature. The solution was stirred at ambienttemperature for 18 h, and was then concentrated to giveN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxypiperidine-4-carboxamidehydrochloride (250 mg) as a white solid that was used without furtherpurification. MS (ES+) C₁₆H₂₂ClN₅O₂ requires: 351 found: 316 [M+H]⁺.

Step 3: SynthesisN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-(6-bromo-4-methylpyridin-2-yl)-4-methoxypiperidine-4-carboxamide

A mixture of N-((6-(IH-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxypiperidine-4-carboxamidehydrochloride (280 mg, crude), 2-bromo-6-fluoro-4-methylpyridine (181mg, 0.96 mmol) and DIPEA (373 mg, 2.88 mmol) in IPA (20 mL) was stirredat 100° C. for 60 h. The reaction mixture was then cooled to ambienttemperature and concentrated. The residue was purified by flash columnchromatography on silica gel (isocratic elution, 1:15 methanol-DCM) togive theN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-(6-bromo-4-methylpyridin-2-yl)-4-methoxypiperidine-4-carboxamide(180 mg, yield 47%) as a yellow solid. MS (ES+) C₂₂H₂₅BrN₆O₂ requires:484, 486 found: 485, 487 [M+H]⁺.

Step 4: SynthesisN-((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxy-1-(4-methyl-6-(3-methyl-1H-pyrazol-5-ylamino)pyridin-2-yl)piperidine-4-carboxamide

A mixture ofN-((6-(H-pyrazol-1-yl)pyridin-3-yl)methyl)-1-(6-bromo-4-methylpyridin-2-yl)-4-methoxypiperidine-4-carboxamide(80 mg, 0.164 mmol), 3-methyl-1H-pyrazol-5-amine (47.7 mg, 0.492 mmol),Pd₂(dba)₃ (30 mg, 0.033 mmol), t-Bu-XPhos (27.8 mg, 0.0656 mmol) andKOAc (64.1 mg, 0.655 mmol) in toluene (2 mL) was stirred at 120° C.under microwave irradiation for 2 h. The reaction mixture was thencooled to ambient temperature, filtered, and concentrated. The residuewas purified by flash column chromatography on silica gel (isocraticelution, 1:30 methanol-DCM). The compound was then further purified byreverse-phase HPLC (ACN/H2O gradient elution with 0.1% TFA) to give thetitle compound (30 mg, yield 37%) as a tan solid.

Example 6. Synthesis of Compound 194 Step 1: Synthesis of1-(6-bromo-4-methylpyridin-2-yl)-3-ethoxyazetidine-3-carboxylic acid

A mixture of 2-bromo-6-fluoro-4-methylpyridine (493 mg, 2.59 mmol),3-ethoxyazetidine-3-carboxylic acid hydrochloride (471 mg, 2.59 mmol),DIPEA (3.17 mL, 18.2 mmol), in ethanol (4.30 mL) was heated to 100° C.for 18 h. The reaction mixture was then concentrated and used in thenext step without any further purification. MS (ES+) C₁₂H₁₅BrN₂O₃requires: 314 found: 315 [M+H]⁺.

Step 2: Synthesis of(S)-1-(6-bromo-4-methylpyridin-2-yl)-3-ethoxy-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)azetidine-3-carboxamide

HATU (855 mg, 2.25 mmol) was added to a mixture of1-(6-bromo-4-methylpyridin-2-yl)-3-ethoxyazetidine-3-carboxylic acid(590 mg, 1.88 mmol),(S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethanamine hydrochloride(550 mg, 2.27 mmol), and DIPEA (1.96 mL, 11.3 mmol) in DMF (4.7 mL) at20° C. The reaction mixture was stirred for 5 min, and then was treatedwith aqueous sodium hydroxide solution (5 M, 20 mL). After stirring foran additional 20 min, the mixture was filtered, and the solid waspurified by flash-column chromatography on silica gel (gradient elution,0 to 10% methanol-DCM) to give(S)-1-(6-bromo-4-methylpyridin-2-yl)-3-ethoxy-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)azetidine-3-carboxamide(980 mg, 100%) as a yellow solid. MS (ES+) C₂₂H₂₄BrFN₆O₂ requires: 502found: 3503 [M+H]⁺.

Step 3: Synthesis of(S)-3-ethoxy-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-1-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)azetidine-3-carboxamide

A mixture of(S)-1-(6-bromo-4-methylpyridin-2-yl)-3-ethoxy-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)azetidine-3-carboxamide(432 mg, 0.824 mmol), 3-methyl-1H-pyrazol-5-amine (280 mg, 2.88 mmol),Pd₂(dba)₃ (38 mg, 0.041 mmol), t-Bu-XPhos (70 mg, 0.17 mmol) and sodiumtert-butoxide (317 mg, 3.30 mmol) in toluene (2.7 mL) was stirred at115° C. for 15 min. The reaction mixture was then cooled to ambienttemperature, loaded onto silica gel, and purified by flash columnchromatography on silica gel (gradient elution, 0 to 10% methanol-DCM)to give the title compound (94 mg, 21%) as a light-pink solid.

Example 7. Synthesis of Heterocycle Intermediates A. Ethyl3-methoxy-2-methylazetidine-3-carboxylate

Step 1: Synthesis of1-(tert-butoxycarbonyl)-3-methoxy-2-methylazetidine-3-carboxylic acid

To a mixture of tert-butyl 2-methyl-3-oxoazetidine-1-carboxylate (360mg, 1.94 mmol), bromoform (1.36 mL) in methanol (8 mL) was addedpotassium hydroxide (872 mg, 15.6 mmol) at 0° C. The cooling bath wasremoved, and the reaction mixture was stirred at 20° C. for 16 h. Theresidue was then concentrated to afford1-(tert-butoxycarbonyl)-3-methoxy-2-methylazetidine-3-carboxylic acid(500 mg, crude) as white solid which was used in the next step withoutfurther purification.

Step 2: Synthesis of 1-(tert-butyl) 3-ethyl3-methoxy-2-methylazetidine-1,3-dicarboxylate

To a mixture of afford1-(tert-butoxycarbonyl)-3-methoxy-2-methylazetidine-3-carboxylic acid(500 mg, crude, 1.94 mmol) in DMF (5 mL) was added iodoethane (0.400 mL)in one portion at 0° C. under nitrogen. The mixture was stirred at 25°C. for 16 h. The reaction mixture was partitioned between water (20 mL)and EtOAc (30 mL). The organic phase was separated, washed with water(20 mL), dried over sodium sulfate, filtered, and concentrated. Theresidue was purified by column chromatography on silica gel (gradientelution, 5 to 20% ethyl acetate-petroleum ether) to afford1-(tert-butyl) 3-ethyl 3-methoxy-2-methylazetidine-1,3-dicarboxylate(300 mg) as yellow oil.

Step 3: Synthesis of ethyl 3-methoxy-2-methylazetidine-3-carboxylate

To a solution of 1-(tert-butyl) 3-ethyl3-methoxy-2-methylazetidine-1,3-dicarboxylate (300 mg, 1.10 mmol) inCHCl₃ (10 mL) was added TMSI (440 mg, 2.20 mmol). The mixture wasstirred at 25° C. for 1 h. The reaction mixture was then concentrated togive a yellow oil which was used directly in the synthetic protocols 1or 2 as a heterocycle intermediate.

B. Ethyl 3-hydroxypiperidine-4-carboxylate

Step 1: Synthesis of ethyl 1-benzyl-3-hydroxypiperidine-4-carboxylate

Ethyl 1-benzyl-3-oxopiperidine-4-carboxylate hydrochloride (1.06 g, 3.56mmol) in MeOH (18 mL) was treated with sodium borohydride (0.457 g, 12.1mmol) at 20° C. The reaction mixture was stirred for 20 min, and thenadditional sodium borohydride (0.220 g, 5.82 mmol) was added. Afterstirring for 20 min, a third portion of sodium borohydride (0.054 g, 1.4mmol) was added, and the resulting mixture stirred for an additional 20min. Water was then added to the reaction mixture, and the MeOH removedin vacuo. The aqueous mixture was extracted with ethyl acetate (3×), andthe combined organic extracts were washed with brine. The washed organiclayer was dried over sodium sulfate, filtered and concentrated in vacuoto give a mixture of ethyl and methyl1-benzyl-3-hydroxypiperidine-4-carboxylate (0.67 g). Ethyl ester: MS(ES+) C₁₅H₂₁NO₃ requires: 263, found: 264 [M+H]⁺. Methyl ester: MS (ES+)C₁₄H₂₉NO₃ requires: 249, found: 250 [M+H]⁺.

Step 2: Synthesis of Ethyl 3-hydroxypiperidine-4-carboxylate

A mixture of ethyl and methyl 1-benzyl-3-hydroxypiperidine-4-carboxylate(0.667 g) and Pd(OH)₂/C (50 mg, 0.071 mmol) were stirred in MeOH (15 mL)under a balloon of H₂ at 20° C. for 18 hours. The reaction mixture wasfiltered through celite, rinsing with methanol, and the filtrate wasconcentrated to give a mixture of ethyl and methyl3-hydroxypiperidine-4-carboxylate (0.405 g) as a pale yellow gum. Thecrude product was used directly in synthetic protocols 1 or 2 as aheterocycle intermediate.

Example & Synthesis of Amine Intermediates A.(S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-amine

Step 1: 1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-one

4-Fluoro-1H-pyrazole (4.73 g, 55 mmol) and potassium carbonate (17.27 g,125 mmol) were combined and stirred in N,N-dimethylformamide (41.7 mL)for 10 minutes in an open sealed tube before addition of2-bromo-5-acetylpyridine (10 g, 50 mmol). The reaction tube was sealedand stirred 20 hours at 100° C. The reaction mixture was then cooled toroom temperature and poured into water (˜700 mL). The mixture wassonicated and stirred for 20 minutes. A beige solid was isolated byfiltration, washed with small amounts of water, and dried to yield1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-one (9.81 g, 96%yield). MS: M+1=206.0.

Step 2:(R)—N—((S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide

To a stirred room temperature solution of1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-one (9.806 g, 47.8mmol) in THF (96 mL) was added (R)-(−)-t-Butylsulfinamide (5.79 g, 47.8mmol) followed by titanium (IV) ethoxide (21.8 g, 96 mmol). The solutionwas stirred at 75° C. on an oil bath for 15 hours. The reaction solutionwas cooled to room temperature and then to −78° C. (externaltemperature) before the next step. To the −78° C. solution was addeddropwise over nearly 55 minutes L-Selectride (143 mL of 1N in THF, 143mmol). During addition, some bubbling was observed. The reaction wasthen stirred after the addition was completed for 15 minutes at −78° C.before warming to room temperature. LC-MS of sample taken during removalfrom cold bath showed reaction was completed. The reaction was cooled to−50° C. and quenched slowly with methanol (˜10 mL), then poured intowater (600 mL) and stirred. An off-white precipitate was removed byfiltration, with ethyl acetate used for washes. The filtrate was dilutedwith ethyl acetate (800 mL), the layers were separated, and the organiclayer was dried over sodium sulfate, filtered, and concentrated down.The crude was purified by silica gel chromatography to yield(R)—N—((S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide(10.5 g, 99% purity, 70.3% yield) as a light yellow solid. MS:M+1=311.1.

Step 3: (S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-amine

A solution of(R)—N—((S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide(10.53 g, 33.9 mmol)) in methanol (79 mmol) and 4N HCl/dioxane (85 mL,339 mmol) was stirred 2.5 hours. LC-MS showed reaction was completed.The reaction solution was poured into diethyl ether (300 mL). A stickysolid was formed. The mixture was treated with ethyl acetate (200 mL)and sonicated. The solvents were decanted, and the sticky solid wastreated with more ethyl acetate (˜200 mL), sonicated and stirred. Thebulk of the sticky solid was converted to a suspension. A light yellowsolid was isolated by filtration, washed with smaller amounts of ethylacetate, and dried to yield(S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethan-1-amine (7.419 g,78% yield). LC-MS confirmed desired product in high purity. MS:M+1=207.1.

B. (S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-anine

Step 1: 1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-one

Sodium hydride (60 wt. %, 276 mg, 6.90 mmol) was added to a mixture of1-(5-chloropyrazin-2-yl)ethanone (800 mg, 5.11 mmol) and4-Fluoro-1H-pyrazole (484 mg, 5.62 mmol) in N,N-dimethylformamide (6.0mL) at ambient temperature for 10 minutes. The reaction mixture was thenpoured into water (70 mL) and was sonicated and stirred for 20 minutes.A dark red solid was isolated by filtration, washed with small amountsof water, and dried to1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-one (919 mg, 95%yield). MS: M+1=207.

Step 2:(R)—N—((S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethyl)-2-methylpropane-2-sulfinamide

To a stirred room temperature solution of1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-one (4.67 g, 22.7mmol) in THF (45 mL) was added (R)-(−)-t-Butylsulfinamide (2.75 g, 22.7mmol) followed by titanium (IV) ethoxide (10.3 g, 45.3 mmol). Thesolution was stirred at 75° C. on an oil bath for 20 hours. The reactionsolution was cooled to room temperature and then to −78° C. before thenext step. To the −78° C. solution was added dropwise over 50 minutesL-Selectride (50.1 mL of 1 N in THF, 50.1 mmol). During addition, somebubbling was observed. The reaction was then stirred after the additionwas completed for 15 minutes at −78° C. before warming to roomtemperature. LC-MS of sample taken during removal from cold bath showedreaction was completed. The reaction was cooled to −60° C. and quenchedslowly with methanol (1 mL), then poured into water (100 mL) andstirred. The mixture was filtered and the solids were washed furtherwith ethyl acetate. The filtrate was diluted with ethyl acetate, thelayers were separated, and the organic layer was dried over sodiumsulfate. The dried solution was filtered, and the filtrate wasconcentrated. The residue was purified by flash-column chromatography(gradient elution, 0 to 100% ethyl acetate-dichloromethane) to give(R)—N—((S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethyl)-2-methylpropane-2-sulfinamide(1.04 g, 14%) as a brown solid. MS: M+1=312. ¹H NMR (400 MHz, DMSO-d6) δ9.12 (d, J=1.4 Hz, 1H), 8.73 (d, J⁼4.5 Hz, 1H), 8.59 (d, J=1.4 Hz, 1H),8.03 (d, J=4.1 Hz, 1H), 5.69 (d, J=5.7 Hz, 1H), 4.62 (p, J=6.8 Hz, 3H),1.57 (d, J=6.9 Hz, 3H), 1.12 (s, 9H).

Step 3: (S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-amine

A solution of(R)—N—((S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethyl)-2-methylpropane-2-sulfinamide(1.04 g, 3.34 mmol) in methanol (7.8 mL) and 4N HCl/dioxane (8.34 mL,33.4 mmol) was stirred for 1.5 h at ambient temperature. The reactionmixture was poured into diethyl ether (100 mL). A light beige solid wasisolated by filtration to afford(S)-1-(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)ethan-1-amine (689 mg,85% yield). MS: M+1=208.

C. (5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)methanamine

Step 1: 5-(4-fluoro-1H-pyrazol-1-yl)pyrazine-2-carbonitrile

To a solution of 5-chloropyrazine-2-carbonitrile (280 mg, 2.0 mmol) inDMF was added 4-fluoro-1H-pyrazole (170 mg, 2.0 mmol), and potassiumacetate (395 mg, 4.0 mmol). The mixture was stirred at the 100° C. for 4hours. The reaction mixture was cooled to 20° C., poured into brine (25mL), and extracted with ethyl acetate. The organic layer was dried oversodium sulfate, concentrated and purified by column chromatography(hexane:ethyl acetate=5:1) to give5-(4-fluoro-1H-pyrazol-1-yl)pyrazine-2-carbonitrile (310 mg, Yield 82%).The structure was confirmed by LC-MS.

Step 2: (5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)methanamine

A mixture of 5-(4-fluoro-1H-pyrazol-1-yl)pyrazine-2-carbonitrile (190mg, 1.0 mmol) and NiCl₂ (12 mg, 0.1 mmol) in MeOH (5 mL) was added NaBH₄(380 mg, 10 mmol) at 0° C. The mixture was stirred at 0° C. for 2 hours,quenched with aqueous NH₄Cl and purified by HPLC to give(5-(4-fluoro-1H-pyrazol-1-yl)pyrazin-2-yl)methanamine (160 mg, Yield82%). The structure was confirmed by LC-MS.

D. (6-(4-fluoro-1H-pyrazol-1-yl)pyridazin-3-yl) methanamine

Step 1: Synthesis of tert-butyl((6-chloropyridazin-3-yl)methyl)carbamate

To a solution of (6-chloropyridazin-3-yl)methanamine (300 mg, 2.0 mmol)in a mixture of THF (9 mL) and water (9 mL), was added NaHCO₃ (200 mg,2.4 mmol) and (Boc)₂O (570 mg, 2.6 mmol). The mixture was stirred at 20°C. for 2 h, and then the reaction mixture was concentrated to remove theTHF. The water was extracted with DCM (2×), and the combined organiclayers were dried over sodium sulfate, filtered, and concentrated togive The organic solvent was dried with anhydrous Na₂SO₄, concentratedby reduce pressure to give tert-butyl((6-chloropyridazin-3-yl)methyl)carbamate (460 mg).

Step 2: Synthesis tert-butyl((6-(4-fluoro-1H-pyrazol-1-yl)pyridazin-3-yl)methyl)carbamate

A mixture of tert-butyl ((6-chloropyridazin-3-yl)methyl)carbamate (300mg, 1.2 mmol), 4-fluoro-1H-pyrazole (106 mg, 1.2 mmol) and Cs₂CO₃ (1.2g, 3.6 mmol) in ACN (20 ml) was stirred at 80° C. for 6 h. The reactionmixture was then filtered, and the filtrate was concentrated to givetert-butyl ((6-(4-fluoro-1H-pyrazol-1-yl)pyridazin-3-yl)methyl)carbamate(246 mg, Purity: 80%), which was used in the next step without furtherpurification.

Step 3: Synthesis(6-(4-fluoro-1H-pyrazol-1-yl)pyridazin-3-yl)methanamine

A solution of tert-butyl((6-(4-fluoro-1H-pyrazol-1-yl)pyridazin-3-yl)methyl)carbamate (246 mg,0.9 mmol) in HCl (4.0 M in dioxane, 10 mL) was stirred at 20° C. for 2h. The reaction mixture was then concentrated to give the title compound(162 mg, Purity: 80%), which was used without further purification. MS(ES+) C₈H₈FN₅ requires: 193, found: 194 [M+H]⁺.

E. (6-(3,5-dimethyl-1H-pyrazol-1-yl)pyridin-3-yl)methanamine

Step 1: 6-(3,5-Dimethyl-1H-pyrazol-1-yl)nicotinonitrile

To the solution of 6-chloronicotinonitrile (300 mg, 2.2 mmol) in DMF (10mL), was added 3,5-dimethyl-1H-pyrazole (210 mg, 2.2 mmol) and Cs₂CO₃(1.4 g, 4.4 mmol). The mixture was stirred at 90° C. for 16 h. Water (25mL) was added to the reaction mixture, and the mixture was filtered. Thesolids were washed with water and dried under vacuum to give6-(3,5-Dimethyl-1H-pyrazol-1-yl)nicotinonitrile (320 mg, yield 74.6%).

Step 2: tert-Butyl((6-(3,5-dimethyl-1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate

To 6-(3,5-Dimethyl-1H-pyrazol-1-yl)nicotinonitrile (300 mg, 1.5 mmol) inMeOH (10 mL), was added NiCl₂ (19 mg, 0.15 mmol), (Boc)₂O (654 mg, 3.0mmol) and NaBH₄ (142 mg, 3.8 mmol). The mixture was stirred at ambienttemperature for 3 h. Saturated aqueous ammonium chloride solution wasadded and the MeOH was removed under vacuum. The aqueous suspension wasthen partitioned with ethyl acetate, and the layers were separated. Theorganic layer was washed with saturated sodium bicarbonate solution(2×50 mL). The organic layer was dried with anhydrous sodium sulfate,filtered, and concentrated under vacuum to give 450 mg target compoundwhich was used in the next step without further purification.

Step 3: 6-(3,5-Dimethyl-1H-pyrazol-1-yl)nicotinonitrile

A solution of HCl in Dioxane (4.0 M, 10 mL) was added to compoundtert-Butyl((6-(3,5-dimethyl-1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate (450mg). The mixture was stirred for 2 h, and then was dried under reducedpressure to give the title compound (350 mg) as a light brown solid thatwas used without further purification. ¹H NMR (400 MHz, DMSO-d6) δ 8.51(d, J=2.1 Hz, 1H), 8.34 (s, 3H), 8.03 (dd, J=8.5, 2.4 Hz, 1H), 7.87 (d,J=8.5 Hz, 1H), 6.14 (s, 1H), 4.12 (q, J=5.7 Hz, 2H), 2.59 (s, 3H), 2.21(s, 3H).

F. (6-(4-chloro-1H-pyrazol-1-yl)pyridin-3-yl)methanamine

Step 1: 6-(4-Chloro-1H-pyrazol-1-yl)nicotinonitrile

To a solution of 6-chloronicotinonitrile (300 mg, 2.2 mmol) in DMF (10mL), was added 4-chloro-1H-pyrazole (227 mg, 2.2 mmol) and Cs₂CO₃ (1.4g, 4.4 mmol). The mixture was stirred at 90° C. for 16 h. Water (25 mL)was added to the mixture, and the mixture was filtered. The solids werewashed with water and dried under vacuum to give6-(4-Chloro-1H-pyrazol-1-yl)nicotinonitrile (380 mg, 84%).

Step 2: tert-Butyl((6-(4-chloro-1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate

To 6-(4-chloro-1H-pyrazol-1-yl)nicotinonitrile (350 mg, 1.7 mmol) inMeOH (10 mL), was added NiCl₂ (19 mg, 0.17 mmol), (Boc)₂O (741 mg, 3.4mmol) and NaBH₄ (163 mg, 4.3 mmol). The mixture was stirred at ambienttemperature for 3 h. Saturated aqueous ammonium chloride solution wasadded and the MeOH was removed under vacuum. The aqueous suspension wasthen partitioned with ethyl acetate, and the layers were separated. Theorganic layer was washed with saturated sodium bicarbonate solution(2×50 mL). The organic layer was dried with anhydrous sodium sulfate,filtered, and concentrated under vacuum to give 480 mg target compoundwhich was used in the next step without further purification.

Step 3: (6-(4-chloro-1H-pyrazol-1-yl)pyridin-3-yl)methanamine

A solution of HCl in Dioxane (4.0 M, 10 mL) was added to tert-Butyl((6-(4-chloro-1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate (450 mg, 1.5mmol) at ambient temperature. The mixture was stirred for 2 h, and thenwas dried under reduced pressure to give the title compound (290 mg) asa light brown solid that was used without further purification. MS:M+1=209.

G. (6-(3-(difluoromethyl)-1H-pyrazol-1-yl)pyridin-3-yl)methanamine

Step 1: 6-(3-formyl-1H-pyrazol-1-yl)nicotinonitrile

To a solution of 1H-pyrazole-3-carbaldehyde (72 mg, 0.75 mmol) and6-chloronicotinonitrile (75 mg, 0.50 mmol) in i-PrOH (2 mL) was addedCs₂CO₃ (100 mg, 0.300 mmol). The resulting mixture was stirred at 100°C. for 2 h. The resulting mixture was then concentrated and crude waspurified by flash-column chromatography on silica gel to give6-(3-formyl-1H-pyrazol-1-yl)nicotinonitrile (1.26 g).

Step 2: 6-(3-(difluoromethyl)-1H-pyrazol-1-yl)nicotinonitrile

To a solution of 6-(3-formyl-1H-pyrazol-1-yl)nicotinonitrile (1.26 g,6.36 mmol) in DCM (120 mL) was added DAST (16.8 mL, 127 mmol) at −78° C.After the addition, the cooling bath was removed and the reactionmixture was stirred for 16 h. The reaction mixture was then treated withsaturated aqueous sodium bicarbonate solution, and the organic layer wasseparated. The organic layer was washed with water, dried over sodiumsulfate, filtered, and concentrated. The residue was purified byflash-column chromatography on silica gel to give6-(3-(difluoromethyl)-1H-pyrazol-1-yl)nicotinonitrile (745 mg).

Steps 3-4:(6-(3-(difluoromethyl)-1H-pyrazol-1-yl)pyridin-3-yl)methanamine

The title compound was prepared from6-(3-(difluoromethyl)-1H-pyrazol-1-yl)nicotinonitrile using the sametwo-step procedure (reduction/protection followed by deprotection) asabove. MS (ES+) C₁₀H₁₀F₂N₄ requires: 224, found: 225 [M+H]⁺.

H. 1-(pyridin-2-yl)pyrrolidin-3-amine

Step 1: Synthesis of tert-butyl(1-(pyridin-2-yl)pyrrolidin-3-yl)carbamate

A mixture of 2-fluoropyridine (1.75 mL, 20.1 mmol), tert-butylpyrrolidin-3-ylcarbamate (250 mg, 1.34 mmol), and DIPEA (0.23 mL, 1.3mmol) was heated to 120° C. for 6 h. The reaction mixture was thenconcentrated to remove excess 2-fluoropyridine, and was then purified byflash-column chromatography on silica gel (gradient elution, 5 to 60%ethyl acetate-hexanes) to give tert-butyl(1-(pyridin-2-yl)pyrrolidin-3-yl)carbamate (180 mg, 51%) as a whitesolid. MS (ES+) C₁₄H₂₁N₃O₂ requires: 263, found: 264 [M+H]⁺.

Step 2: Synthesis of 1-(pyridin-2-yl)pyrrolidin-3-amine

A solution of tert-butyl (1-(pyridin-2-yl)pyrrolidin-3-yl)carbamate (180mg, 0.684 mmol) in ethyl acetate (4 mL) was treated with HCl (4.0 M indioxane, 3.4 mL, 14 mmol) at 20° C. After stirring for 16 h, thereaction mixture was concentrated to give1-(pyridin-2-yl)pyrrolidin-3-amine hydrochloride (162 mg, 100%) as anoff white solid that was used without any further purification. MS (ES+)C₉H₁₃N₃ requires: 163, found: 164 [M+H]⁺.

I. 1-(6-(1H-pyrazol-1-yl)pyridin-3-yl)-N-methylmethanamine

Step 1: Synthesis of tert-butyl((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate

To a solution of (6-(1H-pyrazol-1-yl)pyridin-3-yl)methanamine (1.0 g,5.8 mmol) in DCM (20 mL) was added TEA (1.75 g, 17.3 mmol) and Boc₂O(1.9 g, 8.6 mmol) at 25° C. The resulting mixture was stirred at 25° C.for 16 h. The reaction mixture was then partitioned between ethylacetate and water. The organic layer was washed with brine, dried oversodium sulfate, filtered, and concentrated. The residue was purified byflash column chromatography on silica gel (gradient elution, 0-10%methanol-DCM) to give tert-butyl((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)carbamate (1.3 g, yield 82%) asa light yellow solid. MS (ES+) C₁₄H₁₈N₄O₂ requires: 274, found 275[M+H]⁺.

Step 2: Synthesis of tert-butyl((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)(methyl)carbamate

To a solution of tert-butyl(6-(1H-pyrazol-1-yl)pyridin-3-yl)methylcarbamate (1.2 g, 4.3 mmol) inTHF (30 mL) was added NaH (347 mg, 8.38 mmol, 60% dispersion in mineraloil) at 0° C. under nitrogen. The resulting mixture was stirred at 0° C.for 30 min, and then MeI (740 mg, 5.21 mmol) was added in one portionand the solution was allowed to stir at 25° C. for 3 hours. The reactionmixture was then partitioned between water and EtOAc. The organic layerwas washed with brine, dried over sodium sulfate, and concentrated. Theresidue was purified by flash column chromatography on silica gel(gradient elution, 0-10% methanol-DCM) to give tert-butyl((6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl)(methyl)carbamate (950 mg,yield 76%) as a white powder. MS (ES+) C₁₅H₂₀N₄O₂ requires: 288, found233 [M+H−56]+.

Step 3: Synthesis of1-(6-(1H-pyrazol-1-yl)pyridin-3-yl)-N-methylmethanamine

To a solution of tert-butyl(6-(1H-pyrazol-1-yl)pyridin-3-yl)methyl(methyl)carbamate (940 mg, 3.26mmol) in MeOH (5 mL) was added HCl (4.0 M in dioxane, 3.0 mL) at 25° C.The resulting mixture was stirred at 25° C. for 16 h. The reactionmixture was then concentrated, and the residue was washed with diethylether to give the hydrochloride salt of title compound (640 mg, yield88%) as a white powder. MS (ES+) C₁₀H₁₂N₄ requires: 188, found: 189[M+H]⁺.

The following amine intermediates were made according to the aboveprocedures:

TABLE 5 MS (M + Starting Material Amine Intermediate 1)

221

182

214

196

196

226

Example 9. Synthesis of Other Compound of the Disclosure

The synthetic protocols depicted above were used to prepare additionalcompounds of the disclosure as indicated below. The NMR and LC MS dataobtained for compounds disclosed herein are also shown below in Table 6.

TABLE 6 Compound Synthetic MS Number Protocol ¹H NMR (M + 1) 100 1 ¹HNMR (400 MHz, DMSO-d6) δ ppm 11.85 (s., 1H), 406 9.17 (br. s., 1H), 8.35(t, 1H, J = 12.4 Hz), 7.41-7.30 (m, 2H), 7.24-7.20 (m, 3H), 6.21-6.11(m, 2H), 4.67 (d, 2H, J = 13.2 Hz), 4.26 (d, 2H, J = 6.0 Hz), 2.82 (t,2H, J = 11.6 Hz), 2.47-2.41 (m, 1H), 2.19 (s, 3H), 2.11 (s, 3H),1.76-1.72 (m, 2H), 1.56-1.46 (m, 2H). 101 1 ¹H-NMR (400 MHz, DMSO-d6) δppm 11.84 (s, 1H), 420 9.14 (s, 1H), 8.22 (t, 1H, J = 6.0 Hz), 7.31-7.17(m, 5H), 6.14-6.09 (br, 2H), 4.29 (d, 2H, J = 6.4 Hz), 4.03-4.00 (m,2H), 3.38-3.27 (m, 2H), 2.17 (s, 3H), 2.08 (s, 3H), 2.04-1.99 (m, 2H),1.37-1.31 (m, 2H), 1.13 (s, 3H). 102 4 ¹H-NMR (400 MHz, DMSO-d6) δ ppm11.61 (s, 1H), 421 8.52 (s, 1H), 8.35 (t, 1H, J = 6.4 Hz), 7.31-7.18 (m,5H), 6.28 (s, 1H), 5.98 (s, 1H), 5.97 (s, 1H), 5.50 (s, 1H), 4.27 (d,2H, J = 6.4 Hz), 4.06-4.03 (m, 2H), 3.15-3.08 (m, 2H), 2.15 (s, 3H),2.10 (s, 3H), 1.94-1.88 (m, 2H), 1.50-1.46 (m, 2H). 103 3 ¹H-NMR (400MHz, DMSO-d6) δ ppm 11.86 (s, 1H), 422 9.21 (s, 1H), 8.37 (t, 1H, J =6.4 Hz), 7.32-7.20 (m, 5H), 6.21-6.10 (m, 2H), 5.52 (s, 1H), 4.48-4.45(m, 2H), 4.27 (d, 2H, J = 6.4 Hz), 3.16 (t, 2H, J = 12.0 Hz), 2.19 (s,3H), 2.11 (s, 3H), 1.91-1.82 (m, 2H), 1.51-1.148 (m, 2H). 104 1 ¹H NMR(400 MHz, DMSO-d6) δ 11.8 s, 1H), (9.31- 436 9.03 (m, 1H), 8.52 (t, J =6.2 Hz, 1H), 7.96 (s, 1H), 7.36-6.99 (m, 5H), 6.15 (m 2H), 4.49-4.13 (m,4H), 3.21-3.04 (m, 5H). 2.18 (s, 3H), 2.10 (s. 3H), 1.78 (dd, J = 7.7,4.1 Hz, 4H). 105 4 ¹H -NMR (400 MHz, CDCl3) δ ppm 7.17 (d, 2H, J = 4518.4 Hz), 6.83 (d, 2H, J = 8.4 Hz), 6.08 (s, 1H), 5.99 (s, 1H), 5.74 (s,1H), 4.36 (d, 2H, J = 5.6 Hz), 4.04-4.00 (m, 2H), 3.77 (s, 3H),3.25-3.22 (m, 2H), 2.20-2.16 (m, 7H), 1.70-1.66 (m, 2H), 1.26-1.24 (m,2H). 106 3 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 11.86 (s, 1H), 452 9.20 (s,1H), 8.26 (t, 1H, J = 5.6 Hz), 7.17 (d, 2H, J = 8.8 Hz), 6.86 (d, 2H, J= 8.8 Hz), 6.22-6.11 (br, 1H), 5.50 (s, 1H), 4.48-4.44 (m, 2H), 4.20 (d,2H, J = 6.0 Hz), 3.74 (s, 3H), 3.18-3.11 (m, 2H), 2.19 (s, 3H), 2.11 (s,3H), 1.85-1.84 (m, 2H), 1.48-1.43 (m, 2H). 107 2 ¹H NMR (400 MHz, DMSO)δ 10.71 (s, 1H), 8.73 (t, J = 458 5.7 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H),8.39 (s, 1H), 7.93-7.86 (m, 2H), 7.82 (d, J = 1.0 Hz, 1H), 6.61- 6.54(m, 1H), 6.23 (s, 1H), 5.94 (s, 1H), 5.89 (s, 1H), 4.40 (d, J = 6.0 Hz,4H), 3.94 (d, J = 8.2 Hz, 2H), 2.26 (d, J = 5.6 Hz, 6H), 1.61 (s, 3H).108 4 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 11.73 (br. s., 458 1H), 8.67 (t,2H, J = 5.6 Hz), 8.60 (d, 1H, J = 2.8 Hz), 8.37 (s, 1H), 7.92-7.86 (m,2H), 7.82 (s, 1H), 6.57 (s, 1H), 6.18-6.10 (m, 2H), 5.60 (s, 1H),4.42-4.32 (m, 2H), 3.68-3.63 (m, 1H), 3.54-3.49 (m, 1H), 3.45-3.28 (m,2H), 3.12-3.09 (m, 1H), 2.18-2.12 (m, 5H), 2.10 (s, 3H). 109 3 1H-NMR(400 MHz, DMSO-d6) δ ppm 11.81 (br. s., 459 1H), 9.29 (br. s., 1H),8.68-8.65 (m, 1H), 8.60 (d, 1H, J = 2.0 Hz), 8.37 (d, 1H, J = 1.2 Hz),7.91-7.86 (m, 2H), 7.82 (d, 1H, J = 1.2 Hz), 6.58-6.57 (m, 1H),6.27-6.12 (m, 2H), 4.42-4.31 (m, 2H), 3.79-3.74 (m, 1H), 3.67- 3.62 (m,1H), 3.53-3.44 (m, 2H), 3.12-3.04 (m, 1H), 2.17 (s, 3H), 2.13-2.04 (m,5H). 110 2 ¹H NMR (400 MHz, DMSO) δ 10.77 (s, 1H), 8.83 (t, J = 460 6.2Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.38 (s, 1H), 7.90 (s, 2H), 7.82 (d,J = 1.1 Hz, 1H), 6.59-6.57 (m, 1H), 6.26 (s, 1H), 5.99 (s, 1H), 5.90 (s,1H), 4.41 (dd, J = 17.6, 7.5 Hz, 4H), 4.13 (s, 2H), 2.27 (d, J = 7.2 Hz,6H). 111 2 ¹H NMR (400 MHz, DMSO) δ 9.14 (s, 1H), 8.61 (d, J = 462 2.2Hz, 1H), 8.39 (s, 1H), 7.91 (d, J = 1.3 Hz, 2H), 7.82 (s, 1H), 6.59-6.57(m, 1H), 6.38 (s, 1H), 6.06 (s, 1H), 5.72 (s, 1H), 4.41 (d, J = 6.0 Hz,2H), 4.39-4.32 (m, 2H), 4.19 (dd, J = 21.5, 9.8 Hz, 2H), 2.19 (s, 3H),2.16 (s, 3H). 112 1 ¹H NMR (400 MHz, DMSO) δ 11.23 (s, 1H), 9.28 (s, 4631H), 8.60 (d, J = 2.2 Hz, 1H), 8.42 (s, 1H), 7.95-7.88 (m, 2H), 7.82 (s,1H), 6.58 (s, 1H), 6.49 (s, 1H), 6.23 (s, 1H), 4.68 (d, J = 11.5 Hz,4H), 4.44 (d, J = 5.8 Hz, 2H), 2.34 (s, 3H), 2.23 (s, 3H). 113 4 ¹H-NMR(400 MHz, CDCl3) δ ppm 7.18 (d, 2H, J = 8.4 465 Hz), 6.84 (d, 2H, J =8.4 Hz), 6.15 (s, 1H), 5.97 (s, 1H), 5.78 (s, 1H), 4.37 (d, 2H, J = 5.2Hz), 4.03-3.98 (m, 4H), 3.31-3.29 (m, 2H), 2.22 (s, 3H), 2.19 (s, 3H),2.18- 1.85 (m, 2H), 1.38 (t, 3H, J = 6.4 Hz), 1.26-1.24 (m, 2H). 114 4¹H-NMR (400 MHz, CDCl3) δ ppm 7.13 (d, 2H, J = 8.8 465 Hz), 7.00 (d, 1H,J = 7.2 Hz), 6.75 (d, 2H, J = 8.8 Hz), 6.74 (s, 1H), 5.98 (s, 1H), 5.93(s, 1H), 5.65 (s, 1H), 4.98-4.94 (m, 1H), 3.99-3.92 (m, 2H), 3.69 (s,3H), 3.15-3.08 (m, 2H), 2.16 (s, 3H), 2.14 (s, 3H), 2.12- 2.04 (m, 2H),1.61-1.52 (m, 2H), 1.38 (d, 2H, J = 6.4 Hz). 115 3 ¹H -NMR (400 MHz,DMSO-d6) δ ppm 11.86 (s, 1H), 466 9.21 (br. s., 1H), 8.26 (t, 1H, J =6.4 Hz), 7.15 (d, 2H, J = 8.8 Hz), 6.85 (d, 2H, J = 8.8 Hz), 6.18-6.10(m, 2H), 5.50 (s, 1H), 4.46 (d, 2H, J = 13.2 Hz), 4.20 (d, 2H, J = 6.0Hz), 3.98 (q, 2H, J = 6.8 Hz), 3.15 (t, 2H, J = 12.0 Hz), 2.19 (s, 3H),2.11 (s, 3H), 1.89-1.82 (m, 2H), 1.50- 1.46 (m, 2H), 1.33 (t, 3H, J =6.8 Hz). 116 3 ¹H -NMR (400 MHz, DMSO-d6) δ ppm 11.84 (s, 1H), 466 9.19(s, 1H), 7.92 (d, 1H, J = 8.4 Hz), 7.25 (d, 2H, J = 8.8 Hz), 6.87 (d,2H, J = 8.8 Hz), 6.21-6.08 (s, 2H), 5.47 (s, 1H), 4.88-4.84 (m, 1H),4.48-4.44 (m, 2H), 3.72 (s, 3H), 3.14-3.11 (m, 2H), 2.18 (s, 3H), 2.10(s, 3H), 1.90-1.76 (m, 2H), 1.49-1.37 (m, 2H), 1.48 (d, 3H, J = 7.2 Hz).117 2 ¹H NMR (400 MHz, DMSO) δ 10.63 (s, 1H), 8.63- 472 8.56 (m, 2H),8.43 (d, J = 1.9 Hz, 1H), 7.93 (dt, J = 17.1, 5.3 Hz, 2H), 7.81 (d, J =1.0 Hz, 1H), 6.62-6.53 (m, 1H), 6.23 (s, 1H), 5.90 (d, J = 9.8 Hz, 2H),5.05 (p, J = 6.9 Hz, 1H), 4.36 (dd, J = 12.2, 8.3 Hz, 2H), 3.96- 3.87(m, 2H), 2.25 (d, J = 3.9 Hz, 6H), 1.60 (s, 3H), 1.46 (d, J = 7.0 Hz,3H). 118 1 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 11.84 (br. s., 473 1H), 9.18(s, 1H), 8.60 (d, 1H, J = 2.4 Hz), 8.46 (t, 1H, J = 6.0 Hz), 8.34 (s,1H), 7.90-7.88 (m, 1H), 7.85-7.81 (m, 2H), 6.57 (t, 1H, J = 1.6 Hz),6.21-6.11 (m, 2H), 4.66 (d, 2H, J = 12.8 Hz), 4.32 (d, 2H, J = 6.0 Hz),2.82 (t, 2H, J = 12.0 Hz), 2.46-2.43 (m, 1H), 2.19 (s, 3H), 2.11 (s,3H), 1.77-1.74 (m, 2H), 1.56-1.46 (m, 2H). 119 4 ¹H NMR (400 MHz,DMSO-d6) δ 11.58 (s, 1H), 8.66 476.2 (d, J = 4.5 Hz, 1H), 8.59 (s, 1H),8.55 (d, J = 7.6 Hz, 1H), 8.39 (d, J = 2.2 Hz, 1H), 7.96-7.84 (m, 3H),6.33 (s, 1H), 6.11 (s, 1H), 5.51 (s, 1H), 5.01 (p, J = 7.0 Hz, 1H), 4.00(q, J = 8.6 Hz, 2H), 3.91 (t, J = 6.8 Hz, 1H), 3.83 (t, J = 6.8 Hz, 1H),3.47 (p, J = 7.2 Hz, 1H), 2.14 (s, 3H), 2.08 (s, 3H), 1.41 (d, J = 7.0Hz, 3H). 120 2 ¹H NMR (400 MHz, DMSO) δ 10.47 (s, 1H), 8.69 (dd, 476 J =10.9, 5.1 Hz, 2H), 8.38 (s, 1H), 7.93 (d, J = 4.2 Hz, 1H), 7.89 (d, J =1.3 Hz, 2H), 6.23 (s, 1H), 5.90 (d, J = 10.2 Hz, 2H), 4.37 (dd, J =16.5, 6.9 Hz, 4H), 3.91 (d, J = 8.1 Hz, 2H), 2.24 (s, 6H), 1.60 (s, 3H).121 3 ¹H NMR (400 MHz, DMSO-d6) δ 11.83 (s, 1H), 9.29 477.2 (s, 1H),8.66 (d, J = 4.5 Hz, 1H), 8.56 (d, J = 7.5 Hz, 1H), 8.40 (d, J = 2.2 Hz,1H), 7.96-7.83 (m, 3H), 6.22 (s, 2H), 5.02 (p, J = 7.0 Hz, 1H),4.24-3.82 (m, 3H), 2.17 (s, 3H), 2.10 (s, 3H), 1.41 (d, J = 7.0 Hz, 3H),1.25 (t, J = 6.3 Hz, 2H). 122 1 ¹H NMR (400 MHz, DMSO) δ 11.18 (s, 1H),9.13 (d, J = 477 8.0 Hz, 1H), 8.60 (d, J = 2.3 Hz, 1H), 8.47 (d, J = 2.1Hz, 1H), 8.01 (dd, J = 8.5, 2.2 Hz, 1H), 7.92 (d, J = 8.5 Hz, 1H), 7.82(d, J = 1.0 Hz, 1H), 6.61-6.54 (m, 1H), 6.48 (s, 1H), 6.21 (s, 1H),5.19-5.11 (m, 1H), 4.60 (ddd, J = 34.7, 20.0, 10.0 Hz, 4H), 2.30 (s,3H), 2.23 (s, 3H), 1.52 (d, J = 7.0 Hz, 3H). 123 1 ¹H NMR (400 MHz,DMSO-d6) δ 9.17 (s, 1H), 8.19 (d, 480 J = 8.5 Hz, 2H), 7.34-7.14 (m,2H), 6.85 (d, J = 8.6 Hz, 2H), 6.15 (m, 2H), 4.90 (p, J = 7.1 Hz, 1H),4.28 (d, J = 12.2 Hz, 2H), 3.70 (s, 3H), 3.12 (m, 5H), 2.18 (s, 3H),2.09 (s, 3H), 1.93-1.66 (m, 4H), 1.36 (d, J = 7.0 Hz, 3H). 124 1 ¹H NMR(400 MHz, DMSO) δ 11.18 (s, 1H), 9.23 (t, J = 481 5.7 Hz, 1H), 8.69 (d,J = 4.4 Hz, 1H), 8.42 (d, J = 1.4 Hz, 1H), 7.96-7.88 (m, 3H), 6.49 (s,1H), 6.22 (s, 1H), 4.67 (dd, J = 18.2, 10.9 Hz, 3H), 4.53 (dd, J = 21.0,10.9 Hz, 2H), 4.44 (d, J = 5.9 Hz, 2H), 2.31 (s, 3H), 2.23 (s, 3H). 1251 ¹H NMR (400 MHz, DMSO) δ 8.84 (t, J = 6.0 Hz, 0H), 482 8.76 (d, J =4.7 Hz, 0H), 8.00 (d, J = 1.8 Hz, 0H), 7.71 (d, J = 8.3 Hz, 0H), 6.66(d, J = 8.8 Hz, 0H), 4.42- 4.37 (m, 13H), 4.29 (s, 1H), 4.12 (s, 3H),3.97 (s, 7H), 3.61 (s, 4H), 3.37 (d, J = 12.2 Hz, 3H), 3.06 (s, 0H),2.81 (s, 4H), 2.29 (s, 3H), 2.24 (s, 3H). 126 1 ¹H NMR (300 MHz, DMSO) δ11.18 (s, 1H), 9.34 (d, J = 484 7.4 Hz, 1H), 8.60 (s, 1H), 8.47 (s, 1H),7.99 (d, J = 8.7 Hz, 1H), 7.92 (d, J = 8.6 Hz, 1H), 7.82 (s, 1H), 6.58(s, 1H), 5.07 (s, 1H), 4.66 (s, 2H), 4.60 (s, 2H), 2.30 (s, 3H), 2.25(s, 3H), 1.52 (d, J = 6.9 Hz, 3H). 127 2 ¹H-NMR (400 MHz, DMSO-d6) δ ppm11.62 (s, 1H), 486 8.60 (d, 1H, J = 2.0 Hz), 8.50 (br. s., 1H),8.37-8.33 (m, 2H), 7.91-7.79 (m, 3H), 6.55 (dd, 1H, J = 2.0, 2.8 Hz),6.31 (s, 1H), 6.01 (s, 1H), 5.94 (s, 1H), 4.36 (d, 2H, J = 5.6 Hz),3.75-3.70 (m, 2H), 3.18-3.12 (m, 2H), 2.17 (s, 3H), 2.09 (s, 3H),2.07-2.02 (m, 2H), 1.44-1.37 (m, 2H), 1.15 (s, 3H). 128 1 ¹H NMR (400MHz, DMSO) δ 9.74 (s, 1H), 9.15 (t, J = 487 5.8 Hz, 1H), 8.61 (d, J =2.3 Hz, 1H), 8.40 (s, 1H), 7.91 (s, 2H), 7.82 (s, 1H), 6.57 (s, 1H),6.41 (s, 1H), 6.23 (s, 1H), 4.61 (dd, J = 20.4, 10.5 Hz, 2H), 4.44 (d, J= 6.2 Hz, 2H), 4.38 (d, J = 10.5 Hz, 2H), 2.23 (s, 3H), 2.20 (s, 3H).129 1 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 9.15 (s, 1H), 487 8.57 (d, 1H, J =2.0 Hz), 8.36-8.32 (m, 2H), 7.88-7.78 (m, 3H), 6.53 (dd, 1H, J = 1.6,2.0 Hz), 6.20-6.08 (m, 2H), 4.33 (d, 2H, J = 5.6 Hz), 4.03-3.99 (m, 2H),3.38- 3.27 (m, 2H), 2.17 (s, 3H), 2.08 (s, 3H), 2.00-1.98 (m, 2H),1.37-1.31 (m, 2H), 1.14 (s, 3H). 130 1 ¹H NMR (301 MHz, DMSO) δ 12.13(s, 1H), 11.07 (s, 487 1H), 8.64 (s, 1H), 8.59 (d, J = 2.4 Hz, 1H), 8.44(s, 1H), 7.96 (dd, J = 8.5, 2.1 Hz, 1H), 7.90 (d, J = 8.4 Hz, 1H), 7.81(s, 1H), 6.56 (d, J = 1.8 Hz, 1H), 6.50 (s, 1H), 6.14 (s, 1H), 5.07 (t,J = 7.1 Hz, 1H), 4.39 (d, J = 7.7 Hz, 2H), 4.01 (s, 2H), 2.51 (d, J =1.6 Hz, 3H), 2.27 (s, 3H), 2.01 (d, J = 7.5 Hz, 2H), 1.47 (d, J = 7.0Hz, 3H), 0.76 (t, J = 6.8 Hz, 3H). 131 2 ¹H-NMR (400 MHz, DMSO-d6) δ ppm11.63 (s, 1H), 488 8.60-8.53 (m, 3H), 8.35 (d, 1H, J = 1.2 Hz),7.90-7.84 (m, 2H), 7.81 (d, 1H, J = 1.6 Hz), 6.57-6.56 (m, 1H), 6.34(br. s., 1H), 6.01 (br. s., 1H), 5.98 (s, 1H), 5.55 (s, 1H), 4.33 (d,2H, J = 6.0 Hz), 4.08-4.04 (m, 2H), 3.16- 3.10 (m, 2H), 2.16 (s, 3H),2.11 (s, 3H), 1.95-1.88 (m, 2H), 1.51-1.47 (m, 2H). 132 2 488 133 2 488134 2 488 135 2 488 136 3 ¹H -NMR (400 MHz, DMSO-d6) δ ppm 11.84 (s,1H), 489 9.18 (br. s., 1H), 8.57 (d, 1H, J = 2.0 Hz), 8.54 (t, 1H, J =6.0 Hz), 8.33 (s, 1H), 7.88-7.83 (m, 2H), 7.80 (s, 1H), 6.55 (s, 1H),6.36-6.03 (m, 1H), 5.54 (s, 1H), 4.49-4.44 (m, 2H,), 4.31 (d, 2H, J =6.0 Hz), 3.14-3.10 (m, 3H), 2.18 (s, 3H), 2.11 (s, 3H), 1.87-1.79 (m,2H), 1.50-1.46 (m, 2H). 137 1 ¹H NMR (300 MHz, DMSO) δ 11.14 (s, 1H),8.97 (s, 489 1H), 8.47 (s, 1H), 8.36 (s, 1H), 7.88 (d, J = 8.1 Hz, 1H),7.82 (d, J = 8.0 Hz, 1H), 6.37 (s, 1H), 4.48 (d, J = 10.1 Hz, 2H), 4.40(s, 2H), 4.33 (d, J = 10.3 Hz, 2H), 3.33 (s, 3H), 2.51 (s, 3H), 2.29 (s,3H), 2.24 (s, 3H). 138 1 ¹H NMR (300 MHz, DMSO) δ 11.15 (s, 1H), 8.98(s, 489 1H), 8.36 (s, 2H), 7.84 (t, J = 6.8 Hz, 2H), 7.62 (s, 1H), 6.49(s, 1H), 6.18 (s, 1H), 4.49 (d, J = 10.1 Hz, 2H), 4.38 (d. J = 5.7 Hz,2H), 4.35-4.28 (m, 2H), 3.32 (s, 3H), 2.31 (s, 3H), 2.23 (s, 3H), 2.10(s, 3H). 139 2 ¹H NMR (400 MHz, DMSO) δ 10.61 (s, 1H), 8.67 (d, 490 J =4.6 Hz, 1H), 8.58 (d, J = 7.6 Hz, 1H), 8.42 (d, J = 2.1 Hz, 1H), 7.93(ddd, J = 20.1, 12.2, 5.3 Hz, 3H), 6.22 (s, 1H), 5.90 (d, J = 8.9 Hz,2H), 5.08-5.02 (m, 1H), 4.36 (dd, J = 12.9, 8.3 Hz, 2H), 3.95-3.89 (m,2H), 2.25 (d, J = 3.9 Hz, 6H), 1.59 (s, 3H), 1.46 (d, J = 7.0 Hz, 3H).140 2 ¹H NMR (400 MHz, DMSO) δ 8.68 (d, J = 4.4 Hz, 1H), 490 8.38 (s,1H), 7.95 (s, 1H), 7.91 (s, 2H), 6.29 (s, 1H), 5.95 (s, 1H), 5.84 (d, J= 10.1 Hz, 2H), 4.29-4.20 (m, 4H), 3.99 (d, J = 6.7 Hz, 1H), 2.67 (s,3H), 2.22 (d, J = 2.3 Hz, 6H), 1.53 (d, J = 7.1 Hz, 3H). 141 1 ¹H NMR(400 MHz, DMSO-d6) δ 11.90 (s, 1H), 9.32 491 (s, 1H), 8.90 (q, J = 5.8Hz, 1H), 8.58 (dd, J = 2.6, 0.7 Hz, 1H), 8.34 (d, J = 2.1 Hz, 1H),7.96-7.63 (m, 3H), 6.55 (dd, J = 2.6, 1.7 Hz, 1H), 6.17 (m, 2H), 4.55(d, J = 13.2 Hz, 2H), 4.35 (d, J = 6.1 Hz, 2H), 3.22-2.99 (m, 2H), 2.18(s, 3H), 2.14 (s, 3H), 2.08-1.65 (m, 4H). 142 1 ¹H NMR (400 MHz, DMSO) δ12.19 (s, 1H), 11.10 (s, 491 1H), 8.68 (t, J = 5.8 Hz, 1H), 8.41 (d, J =15.0 Hz, 1H), 7.94 (ddd, J = 14.7, 10.2, 5.9 Hz, 3H), 6.51 (s, 1H), 6.16(s, 1H), 5.91-5.82 (m, 1H), 4.52-4.33 (m, 4H), 3.98 (s, 1H), 2.67 (d, J= 11.6 Hz, 3H), 2.29 (s, 3H), 2.24 (s, 3H), 1.59 (dd, J = 32.0, 6.9 Hz,3H). 143 1 ¹H NMR (400 MHz, CDCl3): δ ppm 8.37-8.48 (m, 1 491 H), 7.94(d, J = 8.8 Hz, 1 H), 7.83 (dd, J = 8.4, 2.4 Hz, 1 H), 7.60 (d, J = 4.4Hz, 1 H), 6.87 (s, 1 H), 6.76 (d, J = 8.0 Hz, 1 H), 6.12 (s, 1 H), 5.99(s, 1 H), 5.29-5.3 (m, 1 H), 4.65 (d, J = 9.2 Hz, 1 H), 4.40-4.41 (m, 1H), 4.04 (d, J = 9.2 Hz, 1 H), 3.36 (s, 3 H), 2.30 (s, 3 H), 2.26 (s, 3H), 1.63 (d, J = 7.2 Hz, 3 H), 1.27 (d, J = 6.8 Hz, 3 H). 144 1 ¹H NMR(400 MHz, CD3OD): δ ppm 8.51 (d, J = 4.0 491 Hz, 1H), 8.41 (s, 1H), 7.92(s, 2H), 7.70 (d, J = 3.6 Hz, 1H), 6.21 (s, 2H), 5.12-5.10 (m, 1H),4.63-4.43 (m, 1H), 4.37-4.25 (m, 1H), 4.16-4.15 (m, 1H), 3.16-3.15 (m,1H), 2.27 (s, 6H), 1.68-1.49 (m, 6H). 145 2 ¹H NMR (400 MHz, DMSO) δ10.49 (s, 1H), 8.65 (dd, 492 J = 21.4, 6.4 Hz, 2H), 8.45 (d, J = 2.1 Hz,1H), 8.01 (dd, J = 8.5, 2.3 Hz, 1H), 7.91 (dd, J = 16.9, 6.4 Hz, 2H),7.14 (t, J = 51.1 Hz, 0H), 6.26 (s, 1H), 5.92 (d, J = 3.5 Hz, 2H), 5.08(dd, J = 14.9, 7.3 Hz, 1H), 4.41 (d, J = 8.6 Hz, 1H), 4.31 (d, J = 8.6Hz, 1H), 4.09-4.02 (m, 3H), 2.24 (d, J = 1.4 Hz, 6H), 1.50 (d, J = 7.1Hz, 3H). 146 1 ¹H NMR (301 MHz, DMSO) δ 9.31 (s, 1H), 8.83 (s, 492 1H),8.60 (d, J = 2.6 Hz, 1H), 8.38 (s, 1H), 7.90 (s, 2H), 7.81 (s, 1H), 6.57(d, J = 1.7 Hz, 1H), 6.32 (d, J = 3.3 Hz, 1H), 6.10 (s, 1H), 4.40 (s,3H), 4.32 (s, 1H), 4.10 (d, J = 9.1 Hz, 2H), 3.30 (s, 3H), 2.22 (s, 3H),2.13 (s, 3H). 147 3 ¹H NMR (400 MHz, DMSO-d6) δ 11.89 (s, 1H), 9.29492.2 (s, 1H), 8.16 (d, J = 7.2 Hz, 1H), 8.03 (dd, J = 5.0, 1.9 Hz, 1H),7.48 (t, J = 8.1 Hz, 1H), 6.61-6.49 (m, 1H), 6.44 (d, J = 8.6 Hz, 1H),6.21 (br. s, 1H), 6.11 (br. s, 1H), 4.40 (q, J = 6.5 Hz, 1H), 4.36-4.21(m, 2H), 3.62 (dd, J = 10.6, 6.8 Hz, 1H), 3.57-3.44 (m, 1H), 3.22-3.10(m, 5H), 2.19 (s, 3H), 2.13 (s, 4H), 1.99 (dd, J = 12.8, 6.6 Hz, 1H),1.80 (br. s, 4H). 148 1 ¹H NMR (301 MHz, DMSO) δ 11.13 (s, 1H), 8.98 (t,J = 493 6.0 Hz, 1H), 8.68 (d, J = 4.6 Hz, 1H), 8.39 (s, 1H), 7.91 (dt, J= 10.9, 6.7 Hz, 3H), 6.49 (s. 1H), 6.17 (s, 1H), 4.46 (d, J = 10.2 Hz,2H), 4.41 (d, J = 5.9 Hz, 2H), 4.32 (d, J = 10.4 Hz, 2H), 3.33 (s, 3H),2.29 (s, 3H), 2.23 (s, 3H). 149 1 ¹H NMR (400 MHz, DMSO) δ 11.22 (s,1H), 8.89 (d, J = 493 7.9 Hz, 1H), 8.67 (d, J = 4.4 Hz, 1H), 8.45 (s,1H), 8.00 (d, J = 8.5 Hz, 1H), 7.91 (dd, J = 12.6, 6.3 Hz, 3H), 6.38 (s,4H), 5.15-5.06 (m, 1H), 4.47 (s, 1H), 4.30 (d, J = 9.2 Hz, 2H), 3.29 (s,3H), 2.24 (s, 3H), 1.50 (d, J = 6.9 Hz, 3H). 150 1 ¹H NMR (400 MHz,DMSO) δ 12.39 (s, 1H), 11.14 (s, 494 1H), 9.06 (t, J = 6.0 Hz, 1H), 8.93(d, J = 4.4 Hz, 1H), 8.21 (d, J = 9.1 Hz, 1H), 8.09 (d, J = 4.1 Hz, 1H),7.86 (d, J = 9.1 Hz, 1H), 6.51 (s, 1H), 6.18 (s, 1H), 4.69 (d, J = 5.9Hz, 2H), 4.51 (d, J = 10.0 Hz, 2H), 4.36 (d, J = 9.8 Hz, 2H), 3.39 (s,3H), 2.30 (s, 3H), 2.25 (s, 3H). 151 2 ¹H NMR (400 MHz, DMSO) δ 9.42 (s,1H), 9.05 (d, J = 494 8.1 Hz, 1H), 8.68 (d, J = 4.5 Hz, 1H), 8.45 (d, J= 2.0 Hz, 1H), 8.00 (dd, J = 8.6, 2.2 Hz, 1H), 7.93 (d, J = 4.3 Hz, 1H),7.90 (d, J = 8.5 Hz, 1H), 6.34 (s, 1H), 6.05 (s, 1H), 5.75 (s, 1H),5.16-5.10 (m, 1H), 4.44-4.30 (m, 2H), 4.24-4.15 (m, 2H), 2.20 (s, 3H),2.16 (s, 3H), 1.50 (d, J = 7.1 Hz, 3H). 152 1 ¹H NMR (400 MHz, DMSO) δ8.84 (s, 1H), 8.68 (d, J = 494 4.5 Hz, 1H), 8.58 (d, J = 7.4 Hz, 1H),8.41 (d, J = 2.1 Hz, 1H), 7.93-7.92 (m, 1H), 7.89 (d, J = 8.4 Hz, 3H),6.36 (s, 1H), 6.04 (s, 1H), 5.36-5.28 (m, 1H), 5.05- 5.00 (m, 1H), 4.12(dd, J = 16.5, 8.2 Hz, 4H), 2.17 (s, 3H), 2.10 (s, 4H), 2.00 (s, 1H),1.43 (d, J = 7.0 Hz, 4H). 153 1 ¹H NMR (400 MHz, DMSO) δ 11.17 (s, 1H),9.13 (d, J = 495 8.1 Hz, 1H), 8.68 (d, J = 4.5 Hz, 1H), 8.47 (d, J = 2.0Hz, 1H), 8.02 (dd, J = 8.5, 2.2 Hz, 1H), 7.92 (dd, J = 13.0, 6.4 Hz,2H), 6.48 (s, 1H), 6.21 (s, 1H), 5.17-5.12 (m, 1H), 4.61 (ddd, J = 31.0,20.8, 9.1 Hz, 4H), 2.30 (s, 3H), 2.23 (s, 3H), 1.51 (d, J = 7.0 Hz, 3H).154 1 ¹H NMR (301 MHz, DMSO) δ 11.13 (s, 1H), 8.91 (s, 496 1H), 7.96 (d,J = 8.3 Hz, 2H), 7.11 (d, J = 10.1 Hz, 1H), 6.48 (s, 1H), 6.18 (s, 1H),5.64 (s, 0H), 5.47 (s, 0H), 4.50-4.17 (m, 6H), 3.95-3.54 (m, 5H), 3.31(s, 3H), 2.26 (d, J = 17.7 Hz, 8H). 155 1 ¹H NMR (301 MHz, DMSO) δ 11.12(s, 1H), 8.91 (t, J = 496 6.0 Hz, 1H), 7.95 (d, J = 7.6 Hz, 2H), 7.10(d, J = 10.0 Hz, 1H), 6.48 (s, 1H), 6.18 (s, 1H), 5.64 (s, 0H), 5.47 (s,0H), 4.51-4.25 (m, 6H), 3.99-3.48 (m, 5H), 3.31 (s, 3H), 2.26 (d, J =17.7 Hz, 8H). 156 1 ¹H NMR (400 MHz, DMSO) δ 9.67 (s, 1H), 8.88 (t, J =499 6.2 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.39 (s, 1H), 7.90 (d, J =1.5 Hz, 2H), 7.82 (d, J = 1.0 Hz, 1H), 6.57 (dd, J = 2.5, 1.7 Hz, 1H),6.36 (s, 1H), 6.23 (s, 1H), 4.44 (s, 1H), 4.41 (d, J = 3.7 Hz, 2H), 4.39(s, 1H), 4.22 (d, J = 9.5 Hz, 2H), 3.31 (s, 3H), 2.22 (s, 3H), 2.20 (s,3H). 157 1 ¹H NMR (400 MHz, DMSO) δ 9.75 (s, 1H), 9.05 (d, J = 501 7.9Hz, 1H), 8.60 (d, J = 2.4 Hz, 1H), 8.46 (d, J = 1.8 Hz, 1H), 8.00 (dd, J= 8.5, 2.2 Hz, 1H), 7.91 (d, J = 8.5 Hz, 1H), 7.82 (s, 1H), 6.58 (d, J =1.8 Hz, 1H), 6.41 (s, 1H), 6.23 (s, 1H), 5.13 (dd, J = 14.5, 7.1 Hz,2H), 4.64- 4.53 (m, 2H), 4.44-4.36 (m, 2H), 2.23 (s, 3H), 2.20 (s, 3H),1.51 (d, J = 7.0 Hz, 3H). 158 1 ¹H NMR (400 MHz, DMSO) δ 11.59 (s, 1H),11.00 (s, 501 1H), 8.60 (d, J = 2.1 Hz, 1H), 8.50 (t, J = 5.7 Hz, 1H),8.39 (s, 1H), 7.90 (d, J = 1.4 Hz, 2H), 7.81 (d, J = 1.0 Hz, 1H), 6.57(dd, J = 2.5, 1.7 Hz, 1H), 6.34 (s, 1H), 6.14 (s, 1H), 4.39 (d, J = 5.7Hz, 2H), 4.19 (s, 2H), 3.23 (t, J = 11.1 Hz, 2H), 2.31 (s, 3H), 2.23 (d,J = 16.4 Hz, 5H), 1.58-1.51 (m, 2H), 1.46 (t, J = 11.0 Hz, 2H), 0.70 (t,J = 7.4 Hz, 3H). 159 1 ¹H NMR (300 MHz, DMSO) δ 11.16 (s, 1H), 9.34 (d,J = 502 7.3 Hz, 1H), 8.69 (d, J = 4.2 Hz, 1H), 8.46 (d, J = 1.9 Hz, 1H),8.06 (dd, J = 8.5, 2.2 Hz, 1H), 7.92 (t, J = 7.1 Hz, 2H), 5.10-5.04 (m,1H), 4.65 (s, 2H), 4.59 (s, 2H), 2.30 (s, 3H), 2.25 (s, 3H), 1.51 (d, J= 6.9 Hz, 3H). 160 4 ¹H-NMR (400 MHz, CDCl3) δ ppm 8.54 (d, 1H, J = 2.0502 Hz), 8.32 (d, 1H, J = 1.6 Hz), 7.90 (d, 1H, J = 8.0 Hz), 7.76 (dd,1H, J = 6.8 Hz, 2.0 Hz), 7.73 (s, 1H), 7.06- 7.02 (m, 2H), 6.47-6.45 (m,1H), 6.14 (br. s., 1H), 5.98 (s, 1H), 5.78 (br. s., 1H), 4.49 (d, 2H, J= 4.4 Hz), 4.00- 3.97 (m, 2H), 3.25 (s, 3H), 3.23-3.18 (m, 2H), 2.25 (s,3H), 2.20 (s, 3H), 2.17-2.11 (m, 2H), 1.89-1.85 (m, 2H). 161 1 ¹H NMR(400 MHz, DMSO-d6) δ 11.83 (s, 1H), 9.18 503 (s, 1H), 8.65 (t, J = 6.2Hz, 1H), 8.58 (d, J = 2.6 Hz, 1H), 8.40-8.22 (m, 1H), 7.97-7.83 (m, 2H),7.79 (d, J = 1.6 Hz, 1H), 6.55 (dd, J = 2.6, 1.7 Hz, 1H), 6.15 (d, J =41.7 Hz, 2H), 4.47-4.19 (m, 4H), 3.23-2.97 (m, 6H), 2.18 (s, 3H), 2.10(s, 3H), 1.89-1.63 (m, 4H). 162 1 ¹H-NMR (400 MHz, CDCl3) δ ppm 8.53 (d,1H, J = 2.4 503 Hz), 8.26 (s, 1H), 7.93 (d, 1H, J = 8.8 Hz), 7.73 (d,1H, J = 8.8 Hz), 7.69-7.67 (m, 1H), 6.98 (s, 1H), 6.47-6.46 (m, 1H),6.05 (s, 1H), 5.98-5.96 (m, 1H), 4.68 (br. s., 2H), 4.62-4.55 (m, 2H),4.12 (br. s., 1H), 3.44-3.37 (m, 2H), 3.11 (br. s., 3H), 2.27 (s, 3H),2.24-2.17 (m, 5H), 1.77-1.74 (m, 2H). 163 1 ¹H NMR (300 MHz, DMSO) δ11.15 (s, 1H), 8.98 (d, J = 503 5.9 Hz, 1H), 8.37 (s, 1H), 7.86 (dd, J =8.5, 2.2 Hz, 1H), 7.77 (d, J = 8.4 Hz, 1H), 6.11 (s, 1H), 4.48 (d, J =10.2 Hz, 2H), 4.40 (d, J = 5.8 Hz, 2H), 4.33 (d, J = 10.0 Hz, 2H), 3.34(s, 3H), 2.56 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H), 2.19 (s, 3H). 164 4¹H NMR (400 MHz, DMSO-d6) δ 11.59 (s, 1H), 8.66 504.2 (d, J = 4.5 Hz,1H), 8.49 (s, 1H), 8.36 (d, J = 8.0 Hz, 2H), 7.95-7.81 (m, 3H), 6.33 (s,1H), 5.96 (d, J = 17.7 Hz, 2H), 4.97 (p, J = 7.0 Hz, 1H), 4.30-4.18 (m,2H), 2.75 (t, J = 12.8 Hz, 2H), 2.41 (td, J = 11.6, 5.8 Hz, 1H), 2.15(s, 3H), 2.09 (s, 3H), 1.75-1.66 (m, 2H), 1.62- 1.43 (m, 2H), 1.39 (d, J= 7.0 Hz, 3H). 165 1 ¹H NMR (400 MHz, DMSO) δ 9.74 (s, 1H), 9.16 (t, J =505 6.1 Hz, 1H), 8.69 (d, J = 4.5 Hz, 1H), 8.40 (d, J = 1.4 Hz, 1H),7.93 (dd, J = 5.5, 3.2 Hz, 2H), 7.89 (d, J = 8.4 Hz, 1H), 6.41 (s, 1H),6.23 (s, 1H), 4.65-4.56 (m, 2H), 4.44 (d, J = 6.0 Hz, 2H), 4.38 (d, J =10.5 Hz, 2H), 2.23 (s, 3H), 2.20 (s, 3H). 166 1 ¹H NMR (300 MHz, DMSO) δ12.16 (s, 1H), 11.07 (s, 505 1H), 8.68 (s, 2H), 8.44 (s, 1H), 7.93 (d, J= 7.4 Hz, 3H), 6.50 (s, 1H), 6.14 (s, 1H), 5.07 (s, 1H), 4.40 (s, 2H),4.01 (s, 2H), 2.51 (s, 3H), 2.26-2.22 (m, 3H), 2.01 (s, 2H), 1.47 (s,3H), 0.76 (s, 3H). 167 5 ¹H NMR (400 MHz, DMSO-d6) δ 11.60 (s, 1H), 8.69506.2 (s, 1H), 8.66 (d, J = 4.6 Hz, 1H), 8.60 (s, 1H), 8.42 (d, J = 2.2Hz, 1H), 7.98 (dd, J = 8.6, 2.3 Hz, 1H), 7.90 (d, J = 4.2 Hz, 1H), 7.87(d, J = 8.5 Hz, 1H), 6.37 (s, 1H), 6.09 (s, 1H), 5.56 (s, 1H), 5.08 (p,J = 7.2 Hz, 1H), 4.08 (dd, J = 41.9, 8.9 Hz, 2H), 3.89 (t, J = 10.8 Hz,2H), 3.25 (s, 3H), 2.15 (s, 3H), 2.08 (s, 3H), 1.47 (d, J = 7.1 Hz, 3H).168 1 ¹H NMR (400 MHz, DMSO) δ 8.91-8.79 (m, 1H), 506 8.72 (d, J = 8.2Hz, 1H), 8.60 (d, J = 2.4 Hz, 1H), 8.44 (d. J = 1.9 Hz, 1H), 7.99 (dd, J= 8.6, 2.2 Hz, 1H), 7.90 (d, J = 8.5 Hz, 1H), 7.81 (s, 1H), 6.58-6.55(m, 1H), 6.41 (s, 1H), 6.02 (s, 1H), 5.13-5.07 (m, 1H), 4.30 (d, J = 8.4Hz, 1H), 4.20 (d, J = 8.9 Hz, 1H), 4.08-4.01 (m, 2H), 3.27 (s, 3H), 2.18(s, 1H), 2.10 (s, 1H), 1.49 (d, J = 7.0 Hz, 1H). 169 1 ¹H NMR (400 MHz,DMSO-d6) δ 9.32 (s, 1H), 8.69- 507.2 8.64 (m, 2H), 8.42 (d, J = 2.2 Hz,1H), 7.98 (dd, J = 8.6, 2.3 Hz, 1H), 7.90 (d, J = 4.3 Hz, 1H), 7.87 (d,J = 8.5 Hz, 1H), 6.28 (s, 1H), 6.18 (s, 1H), 5.08 (p, J = 7.1 Hz, 1H),4.15 (dd, J = 43.2, 9.4 Hz, 2H), 3.97 (t, J = 10.6 Hz, 2H), 3.26 (s,3H), 2.17 (s, 3H), 2.09 (s, 3H), 1.47 (d, J = 7.0 Hz, 3H). 170 2 ¹H NMR(400 MHz, DMSO) δ 8.90 (d, J = 8.1 Hz, 1H), 507 8.68 (d, J = 4.4 Hz,1H), 8.45 (d, J = 1.5 Hz, 1H), 8.00 (d, J = 8.5 Hz, 1H), 7.91 (dd, J =13.0, 6.3 Hz, 2H), 6.20 (d, J = 7.7 Hz, 1H), 5.12-5.08 (m, 7H), 4.47 (d,J = 10.7 Hz, 1H), 4.35 (t, J = 10.1 Hz, 2H), 4.25 (s, 1H), 3.29 (d, J =5.3 Hz, 3H), 2.36 (d, J = 7.1 Hz, 3H), 2.24 (d, J = 4.9 Hz, 3H), 1.50(d, J = 7.0 Hz, 3H). 171 1 ¹H NMR (400 MHz, DMSO) δ 8.90 (s, 1H), 8.69(dd, J = 508 8.6, 4.4 Hz, 1H), 8.39 (d, J = 19.0 Hz, 1H), 7.95 (t, J =3.8 Hz, 1H), 7.90 (d, J = 1.3 Hz, 1H), 6.40 (s, 1H), 6.05 (s, 1H), 5.85(d, J = 7.1 Hz, 1H), 4.28-4.10 (m, 4H), 2.67 (s, 3H), 2.18 (s, 3H), 2.11(s, 3H), 2.01 (d, J = 7.7 Hz, 1H), 1.52 (d, J = 7.1 Hz, 3H). 172 1 ¹HNMR (400 MHz, DMSO) δ 11.13 (s, 1H), 8.98 (t, J = 509 6.1 Hz, 1H), 8.78(s, 1H), 8.41 (d, J = 1.6 Hz, 1H), 7.96 (s, 1H), 7.95-7.92 (m, 1H), 7.89(d, J = 8.4 Hz, 1H), 6.50 (s, 1H), 6.18 (s, 1H), 4.47 (d, J = 9.9 Hz,2H), 4.42 (d, J = 6.0 Hz, 2H), 4.33 (d, J = 9.5 Hz, 2H), 3.33 (s, 3H),2.30 (s, 3H), 2.24 (s, 3H). 173 1 ¹H NMR (400 MHz, DMSO) δ 9.48 (s, 1H),8.85 (t, J = 510 6.1 Hz, 1H), 8.60 (d, J = 2.4 Hz, 1H), 8.38 (s, 1H),7.90 (d, J = 1.2 Hz, 2H), 7.82 (d, J = 1.0 Hz, 1H), 6.86 (t, J = 55.7Hz, 1H), 6.63 (s, 1H), 6.57 (dd, J = 2.4, 1.8 Hz, 1H), 6.15 (s, 1H),5.92 (s, 1H), 4.39 (d, J = 6.1 Hz, 2H), 4.22 (d, J = 9.1 Hz, 2H), 4.05(d, J = 9.1 Hz, 2H), 3.32 (s, 3H), 2.21 (s, 3H). 174 1 ¹H NMR (301 MHz,DMSO) δ 9.04 (s, 1H), 8.83 (s, 510 1H), 8.68 (d, J = 4.4 Hz, 1H), 8.37(s, 1H), 7.94-7.88 (m, 3H), 6.38 (s, 1H), 6.05 (s, 1H), 4.40 (s, 1H),4.29- 4.27 (m, 1H), 4.08 (d, J = 9.1 Hz, 2H), 3.30 (s, 1H), 2.19 (s,3H), 2.11 (s, 3H). 175 1 ¹H NMR (400 MHz, DMSO) δ 9.65 (s, 1H), 8.78 (d,J = 513 8.1 Hz, 1H), 8.60 (d, J = 2.3 Hz, 1H), 8.45 (d, J = 2.1 Hz, 1H),7.99 (dd, J = 8.5, 2.3 Hz, 1H), 7.90 (d, J = 8.5 Hz, 1H), 7.81 (d, J =1.1 Hz, 1H), 6.58-6.55 (m, 1H), 6.35 (s, 1H), 6.22 (s, 1H), 5.09 (dd, J= 15.2, 7.7 Hz, 1H), 4.44 (d, J = 9.5 Hz, 1H), 4.35 (d, J = 9.6 Hz, 1H),4.19 (dd, J = 14.7, 9.5 Hz, 1H), 3.28 (s, 3H), 2.20 (d, J = 4.2 Hz, 6H),1.50 (d, J = 7.0 Hz, 3H). 176 1 ¹H NMR (301 MHz, DMSO) δ 11.11 (s, 1H),8.85 (s, 514 1H), 8.02 (s, 1H), 7.95 (s, 1H), 7.78 (d, J = 8.8 Hz, 1H),6.84 (d, J = 9.2 Hz, 1H), 6.49 (s, 1H), 6.18 (s, 1H), 4.44 (d, J = 10.1Hz, 2H), 4.35-4.24 (m, 4H), 3.94 (t, J = 13.1 Hz, 2H), 3.71 (t, J = 7.3Hz, 2H), 3.30 (s, 3H), 2.63- 2.55 (m, 2H), 2.29 (s, 3H), 2.24 (s, 3H).177 1 ¹H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 8.94 (t, J = 514 6.0 Hz,1H), 8.44 (d, J = 2.5 Hz, 1H), 8.34 (s, 1H), 7.85 (dd, J = 10.8, 8.8 Hz,2H), 6.29-6.25 (m, 2H), 5.95 (s, 2H), 4.37 (dd, J = 12.0, 7.6 Hz, 4H),4.22 (d, J = 9.1 Hz, 2H), 3.31 (s, 3H), 2.25 (s, 6H), 2.01-1.98 (m, 1H),0.97-0.93 (m, 2H), 0.76 (dd, J = 4.8, 2.1 Hz, 2H). 178 1 ¹H NMR (300MHz, DMSO) δ 11.84 (s, 1H), 9.37 (s, 515 1H), 8.78 (t, J = 6.1 Hz, 1H),8.33 (d, J = 3.2 Hz, 2H), 7.83 (t, J = 5.4 Hz, 2H), 7.62 (s, 1H), 6.19(s, 1H), 4.36 (d, J = 6.0 Hz, 2H), 4.20 (d, J = 9.4 Hz, 2H), 4.01 (d, J= 9.4 Hz, 2H), 3.30 (s, 3H), 2.19 (s, 3H), 2.12 (s, 3H), 1.79 (ddd, J =13.4, 8.3, 4.9 Hz, 1H), 0.90-0.84 (m, 2H), 0.63-0.58 (m, 2H). 179 1 ¹HNMR (400 MHz, DMSO) δ 11.94 (s, 1H), 9.64 (s, 515 1H), 8.82 (t, J = 6.1Hz, 1H), 8.44 (d, J = 2.6 Hz, 1H), 8.33 (d, J = 1.7 Hz, 1H), 7.85 (dd, J= 8.5, 2.2 Hz, 1H), 7.80 (d, J = 8.3 Hz, 1H), 6.27 (d, J = 2.6 Hz, 1H),4.37 (d, J = 6.1 Hz, 2H), 4.25 (d, J = 9.4 Hz, 2H), 4.06 (d, J = 9.4 Hz,2H), 3.30 (s, 3H), 2.20 (s, 3H), 2.15 (s, 3H), 2.03-1.96 (m, 1H),0.97-0.92 (m, 2H), 0.78-0.74 (m, 2H). 180 2 ¹H NMR (400 MHz, DMSO) δ9.38 (s, 1H), 9.15 (t, J = 516 6.0 Hz, 1H), 8.69 (d, J = 4.5 Hz, 1H),8.39 (s, 1H), 7.96- 7.87 (m, 3H), 6.87 (s, 2H), 6.73 (s, 1H), 6.13 (s,1H), 5.93 (s, 1H), 4.42 (d, J = 6.5 Hz, 2H), 4.37 (d, J = 10.1 Hz, 1H),4.22 (dd, J = 21.6, 10.1 Hz, 3H), 2.20 (s, 3H). 181 4 ¹H-NMR (400 MHz,DMSO-d6) δ ppm 11.69 (br. s., 516 1H), 8.59-8.50 (m, 3H), 8.42 (d, 1H, J= 1.2 Hz), 7.98 (dd, 1H, J = 6.8 Hz, 1.6 Hz), 7.89 (d, 1H, J = 6.4 Hz),7.80 (s, 1H), 6.56 (d, 1H, J = 1.6 Hz), 6.30 (s, 1H), 5.97 (s, 2H),5.08-5.01 (m, 1H), 3.97-3.89 (m, 2H), 3.15 (s, 3H), 3.12-3.05 (m, 2H),2.16 (s, 3H), 2.10 (s, 3H), 1.86- 1.74 (m, 4H), 1.47 (d, 3H, J = 5.2Hz). 182 2 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 8.56-8.54 (m, 516 2H), 8.30(d, 1H, J = 2.0 Hz), 7.77 (dd, 1H, J = 2.0, 8.4 Hz), 7.72 (d, 1H, J =8.4 Hz), 6.28 (s, 1H), 6.08 (s, 1H), 5.97 (s, 2H), 4.31 (d, 2H, J = 6.0Hz), 4.07-4.03 (m, 2H), 3.16-3.09 (m, 2H), 2.54 (s, 3H), 2.18 (s, 3H),2.15 (s, 3H), 2.10 (s, 3H), 1.94-1.87 (m, 2H), 1.49-1.45 (m, 2H). 183 2¹H NMR (400 MHz, DMSO) δ 9.27 (s, 1H), 9.16 (d, J = 517 5.6 Hz, 2H),8.74 (d, J = 4.5 Hz, 1H), 8.51 (s, 1H), 8.05 (d, J = 4.1 Hz, 1H), 6.80(d, J = 48.2 Hz, 2H), 6.13 (s, 1H), 5.91 (s, 1H), 4.56 (d, J = 5.8 Hz,2H), 4.40 (dd, J = 20.7, 10.2 Hz, 2H), 4.23 (dd, J = 21.4, 9.8 Hz, 2H),2.20 (s, 3H). 184 1 ¹H NMR (400 MHz, DMSO) δ 9.67 (s, 1H), 8.89 (t, J =517 6.1 Hz, 1H), 8.69 (d, J = 3.9 Hz, 1H), 8.38 (s, 1H), 7.91 (dt, J =11.4, 6.3 Hz, 3H), 6.36 (s, 1H), 6.24 (s, 1H), 4.43 (s, 1H), 4.41 (s,2H), 4.39 (s, 1H), 4.23 (s, 1H), 4.20 (s, 1H), 3.31 (s, 3H), 2.21 (s,3H), 2.20 (s, 3H). 185 1 ¹H-NMR (400 MHz, DMSO-d6) δ ppm 11.85 (s, 1H),517 9.22 (s, 1H), 8.61 (d, 1H, J = 2.0 Hz), 8.37 (s, 1H), 7.92-7.85 (m,2H), 7.82 (s, 1H), 6.57 (s, 1H), 6.22-6.12 (m, 2H), 4.58 (s, 2H),4.29-4.26 (m, 2H), 3.28 (s, 3H), 3.27-3.20 (m, 2H), 3.17 (s, 3H), 2.19(s, 3H), 2.11 (s, 3H), 1.99-1.83 (m, 4H). 186 1 ¹H-NMR (400 MHz,DMSO-d6) δ ppm 11.84 (s, 1H), 517 9.19 (br. s., 1H), 8.59 (d, 1H, J =2.0 Hz), 8.50 (d, 1H, J = 8.4 Hz), 8.42 (d, 1H, J = 1.6 Hz), 7.96 (dd,1H, J = 2.0, 8.4 Hz), 7.89 (d, 1H, J = 8.4 Hz), 7.80 (d, 1H, J = 0.8Hz), 6.56-6.55 (m, 1H), 6.20-6.08 (m, 2H), 5.07- 5.03 (m, 1H), 4.36-4.28(m, 2H), 3.18-3.12 (m, 5H), 2.19 (s, 3H), 2.10 (s, 3H), 1.85-1.67 (m,4H), 1.46 (d, 3H, J = 7.2 Hz). 187 1 ¹H-NMR (400 MHz, CDCl3) δ ppm 8.53(d, 1H, J = 2.4 517 Hz), 8.29 (d, 1H, J = 2.0 Hz), 7.94 (d, 1H, J = 8.4Hz), 7.74 (d, 1H, J = 1.2 Hz), 7.69-7.65 (m, 1H), 6.82 (s, 1H), 6.47 (t,1H, J = 2.0 Hz), 6.08-6.06 (m, 2H), 5.98 (s, 1H), 4.61-452 (m, 2H), 4.09(br. s., 1H), 3.44-3.37 (m, 2H), 2.88-2.73 (m, 3H), 2.28 (s, 3H), 2.23(s, 3H), 1.76-1.59 (m, 4H), 1.25 (s, 3H). 188 4 ¹H-NMR (400 MHz, CDCl3)δ ppm 8.36 (d, 1H, J = 4.4 518 Hz), 8.33 (d, 1H, J = 1.2 Hz), 7.86 (d,1H, J = 8.4 Hz), 7.72 (dd, 1H, J = 8.4 Hz, 2.4 Hz), 7.56 (d, 1H, J = 4.4Hz), 7.05 (br. s., 1H), 6.11 (d, 1H, J = 7.2 Hz), 6.04 (br. s., 1H),5.95 (s, 1H), 5.76 (s, 1H), 5.19-5.13 (m, 1H), 3.67-3.60 (m, 2H),3.36-3.31 (m, 2H), 2.24 (s, 3H), 2.16 (s, 3H), 2.06 (br. s., 2H),1.57-1.51 (m, 5H), 1.21 (s, 3H). 189 1 ¹H NMR (400 MHz, DMSO) δ 9.73 (s,1H), 9.05 (d, J = 519 7.6 Hz, 1H), 8.68 (d, J = 4.5 Hz, 1H), 8.45 (d, J= 2.1 Hz, 1H), 8.00 (dd, J = 8.6, 2.2 Hz, 1H), 7.93 (d, J = 4.3 Hz, 1H),7.90 (d, J = 8.5 Hz, 1H), 6.41 (s, 1H), 6.22 (s, 1H), 5.17-5.11 (m, 1H),4.65-4.52 (m, 2H), 4.43- 4.36 (m, 2H), 2.22 (s, 3H), 2.19 (s, 3H), 1.50(d, J = 7.0 Hz, 3H). 190 1 ¹H NMR (400 MHz, DMSO) δ 11.54 (s, 1H), 11.00(s, 519 1H), 8.67 (d, J = 4.3 Hz, 1H), 8.49 (s, 1H), 8.38 (s, 1H),7.94-7.87 (m, 3H), 6.34 (s, 1H), 6.13 (s, 1H), 4.38 (d, J = 4.9 Hz, 2H),4.19 (s, 2H), 3.20 (d, J = 25.1 Hz, 2H), 2.30 (s, 3H), 2.22 (d, J = 17.9Hz, 5H), 1.57-1.41 (m, 3H), 0.69 (t, J = 7.3 Hz, 3H). 191 3 ¹H-NMR (400MHz, CDCl3) δ ppm 8.37 (d, 1H, J = 3.2 519 Hz), 8.35 (d, 1H, J = 1.6Hz), 7.89 (d, 1H, J = 6.8 Hz), 7.74 (dd, 1H, J = 8.4 Hz, 2.0 Hz), 7.57(d, 1H, J = 3.2 Hz), 6.83 (br. s., 1H), 6.05 (s, 1H), 6.00 (s, 1H), 5.88(d, 1H, J = 5.2 Hz), 5.22-5.17 (m, 1H), 4.01-3.97 (m, 2H), 3.64-3.59 (m,2H), 2.30 (s, 3H), 2.25 (s, 3H), 2.04-2.02 (m, 2H), 1.59-1.54 (m, 5H),1.25 (s, 3H). 192 4 ¹H NMR (400 MHz, DMSO-d6) δ 11.60 (s, 1H), 8.66520.2 (d, J = 4.5 Hz, 1H), 8.48 (s, 1H), 8.41 (d, J = 2.2 Hz, 1H), 8.26(d, J = 8.2 Hz, 1H), 7.96 (dd, J = 8.5, 2.3 Hz, 1H), 7.90 (d, J = 4.2Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 6.31 (s, 1H), 5.96 (d, J = 8.3 Hz,2H), 5.48 (s, 1H), 4.99 (p, J = 7.2 Hz, 1H), 4.02 (t, J = 15.5 Hz, 2H),3.11 (t, J = 13.1 Hz, 2H), 2.15 (s, 3H), 2.09 (s, 3H), 1.86 (dtd, J =38.3, 13.0, 4.4 Hz, 2H), 1.61-1.36 (m, 5H). 193 4 ¹H-NMR (400 MHz,DMSO-d6) δ ppm 11.72 (br. s., 520 1H), 8.71-8.67 (m, 2H), 8.59 (s, 1H),8.34 (s, 1H), 7.92 (d, 1H, J = 4.4 Hz), 7.88 (s, 1H), 7.87 (s, 1H), 6.32(s, 1H), 5.98 (s, 2H), 4.34 (d, 2H, J = 6.0 Hz), 3.97-3.94 (m, 2H), 3.16(s, 3H), 3.12-3.05 (m, 2H), 2.16 (s, 3H), 2.11 (s, 3H), 1.88-1.74 (m,4H). 194 5 ¹H NMR (400 MHz, DMSO-d6) δ 11.61 (s, 1H), 8.67 520.2 (d, J =4.5 Hz, 1H), 8.64 (d, J = 8.0 Hz, 2H), 8.42 (d, J = 2.2 Hz, 1H), 7.98(dd, J = 8.5, 2.3 Hz, 1H), 7.91 (d, J = 4.2 Hz, 1H), 7.88 (d, J = 8.5Hz, 1H), 6.38 (s, 1H), 6.09 (s, 1H), 5.56 (s, 1H), 5.08 (p, J = 7.2 Hz,1H), 4.15 (d, J = 8.7 Hz, 2H), 4.04 (d, J = 8.8 Hz, 2H), 3.86 (t, J =9.6 Hz, 2H), 3.44-3.37 (m, 2H), 2.15 (s, 3H), 2.08 (s, 3H), 1.47 (d, J =7.0 Hz, 3H), 1.19 (t, J = 6.9 Hz, 3H). 195 1 ¹H-NMR (400 MHz, DMSO-d6) δppm 11.84 (s, 1H), 521 9.19 (s, 1H), 8.68-8.65 (m, 2H), 8.33 (s, 1H),7.90 (d, 1H, J = 4.0 Hz), 7.86 (s, 1H), 7.85 (s, 1H), 6.27-6.09 (m, 2H),4.36-4.31 (m, 4H), 3.16-3.11 (m, 5H), 2.18 (s, 3H), 2.10 (s, 3H),1.78-1.77 (m, 4H). 196 1 ¹H NMR (301 MHz, DMSO) δ 11.12 (s, 1H), 8.67(d, J = 521 4.5 Hz, 1H), 8.62 (s, 1H), 8.42 (d, J = 1.9 Hz, 1H), 7.98(dd, J = 8.5, 2.2 Hz, 2H), 7.92 (d, J = 4.2 Hz, 1H), 7.89 (s, 1H), 6.36(s, 2H), 5.12-4.99 (m, 1H), 4.13 (s, 1H), 3.37 (s, 2H), 3.17 (s, 3H),2.25 (s, 1H), 1.95 (d, J = 12.8 Hz, 4H), 1.47 (d, J = 7.0 Hz, 3H). 197 1¹H NMR (400 MHz, DMSO-d6) δ 11.86 (s, 1H), 9.36 521.2 (s, 1H), 8.67 (d,J = 4.5 Hz, 1H), 8.64 (d, J = 8.2 Hz, 1H), 8.42 (d, J = 2.3 Hz, 1H),7.97 (dd, J = 8.5, 2.4 Hz, 1H), 7.91 (d, J = 4.2 Hz, 1H), 7.88 (d, J =8.5 Hz, 1H), 6.21 (b.s., 2H), 5.07 (p, J = 7.2 Hz, 1H), 4.29-4.06 (m,2H), 4.03-3.88 (m, 2H), 3.45-3.36 (m, 2H), 2.18 (s, 3H), 2.10 (s, 3H),1.47 (d, J = 7.0 Hz, 3H), 1.19 (t, J = 7.0 Hz, 3H). 198 1 ¹H NMR (400MHz, CDCl3): δ ppm 8.37-8.48 (m, 1 521 H), 7.94 (d, J = 8.8 Hz, 1 H),7.83 (dd, J = 8.4, 2.4 Hz, 1 H), 7.60 (d, J = 4.4 Hz, 1 H), 6.87 (s, 1H), 6.76 (d, J = 8.0 Hz, 1 H), 6.12 (s, 1 H), 5.99 (s, 1 H), 5.29-5.3(m, 1 H), 4.65 (d, J = 9.2 Hz, 1 H), 4.40-4.41 (m, 1 H), 4.04 (d, J =9.2 Hz, 1 H), 3.36 (s, 3 H), 2.30 (s, 3 H), 2.26 (s, 3 H), 1.63 (d, J =7.2 Hz, 3 H), 1.27 (d, J = 6.8 Hz, 3 H). 199 1 ¹H NMR (400 MHz, CDCl3):δ ppm 8.38-8.48 (m, 1 521 H), 7.95 (d, J = 8.4 Hz, 1 H), 7.82 (dd, J =8.4, 2.4 Hz, 1 H), 7.61 (d, J = 4.4 Hz, 1 H), 6.86 (s, 1 H), 6.74 (d, J= 8.4 Hz, 1 H), 6.14 (s, 1 H), 6.03 (s, 1 H), 5.27-5.42 (m, 1 H),4.44-4.65 (m, 1 H), 4.45 (d, J = 6.78 Hz, 1 H), 4.02 (d, J = 9.2 Hz, 1H), 3.26 (s, 3 H), 2.31 (s, 3 H), 2.28 (s, 3 H), 1.62 (d, J = 7.2 Hz, 3H), 1.51 (d, J = 6.8 Hz, 3 H). 200 4 ¹H NMR (400 MHz, DMSO-d6) δ 11.61(s, 1H), 8.78- 522.2 8.70 (m, 1H), 8.66 (d, J = 4.5 Hz, 1H), 8.53 (s,1H), 8.40 (d, J = 2.3 Hz, 1H), 7.96 (dd, J = 8.5, 2.3 Hz, 1H), 7.91 (d,J = 4.3 Hz, 1H), 7.87 (d, J = 8.5 Hz, 1H), 6.37 (s, 1H), 6.00 (s, 1H),5.98 (s, 1H), 5.04 (dt, J = 14.2, 7.0 Hz, 1H), 4.16 (t, J = 16.8 Hz,2H), 3.14-3.00 (m, 2H), 2.15 (s, 3H), 2.10 (s, 3H), 2.01-1.68 (m, 4H),1.46 (d, J = 7.1 Hz, 3H). 201 1 ¹H NMR (400 MHz, DMSO) δ 9.69 (s, 1H),8.75 (d, J = 524 8.2 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.45 (d. J = 2.2Hz, 1H), 7.99 (dd, J = 8.5, 2.3 Hz, 1H), 7.91 (d, J = 8.4 Hz, 1H), 7.81(d, J = 1.0 Hz, 1H), 6.87 (s, 1H), 6.62- 6.54 (m, 2H), 6.15 (s, 1H),5.95 (s, 1H), 5.14-5.07 (m, 1H), 4.27 (d, J = 9.1 Hz, 1H), 4.17 (d, J =9.0 Hz, 1H), 4.05 (dd, J = 13.4, 9.2 Hz, 2H), 2.23 (s, 3H), 1.50 (d, J =7.1 Hz, 3H). 202 1 ¹H NMR (301 MHz, DMSO) δ 8.69 (dd, J = 9.8, 6.4 524Hz, 3H), 8.44 (d, J = 2.0 Hz, 1H), 7.99 (dd, J = 8.5. 2.2 Hz, 1H), 7.90(dd, J = 8.3, 6.5 Hz, 2H), 6.42 (s, 1H), 5.99 (s, 1H), 5.17-5.03 (m,1H), 4.28 (d, J = 8.3 Hz, 1H), 4.18 (d, J = 8.9 Hz, 1H), 4.03 (t, J =8.9 Hz, 2H), 3.26 (s, 3H), 2.16 (s, 1H), 2.09 (s, 1H), 1.48 (d, J = 7.0Hz, 1H). 203 1 ¹H NMR (400 MHz, DMSO) δ 12.29 (s, 1H), 11.14 (s, 5251H), 8.99 (t, J = 6.0 Hz, 1H), 8.90 (s, 1H), 8.44 (s, 1H), 8.09 (s, 1H),7.98-7.92 (m, 2H), 7.14 (s, 1H), 6.50 (s, 1H), 6.18 (s, 1H), 4.47 (d, J= 9.8 Hz, 2H), 4.43 (d, J = 6.0 Hz, 2H), 4.33 (d, J = 10.0 Hz, 2H), 3.34(s, 3H), 2.29 (s, 3H), 2.24 (s, 3H). 204 1 ¹H NMR (400 MHz, DMSO) δ12.24 (s, 1H), 11.14 (s, 525 1H), 8.99 (t, J = 6.1 Hz, 1H), 8.72 (d, J =2.5 Hz, 1H), 8.45 (d, J = 1.7 Hz, 1H), 7.95 (dt, J = 18.2, 5.3 Hz, 2H),7.29 (s, 0H), 7.15 (s, 0H), 7.02 (s, 0H), 6.85 (d, J = 2.6 Hz, 1H), 6.50(s, 1H), 6.18 (s, 1H), 4.45 (dd, J = 15.4, 8.0 Hz, 4H), 4.33 (d, J =10.0 Hz, 2H), 3.34 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H). 205 1 ¹H NMR(400 MHz, DMSO) δ 12.30 (s, 2H), 11.13 (s, 526 1H), 9.14 (d, J = 1.3 Hz,1H), 9.00 (t, J = 5.9 Hz, 1H), 8.74 (d, J = 2.5 Hz, 1H), 8.56 (s, 1H),7.33 (s, 0.2H), 7.19 (s, 0.5H), 7.06 (s, 0.3H), 6.93 (d, J = 2.6 Hz,1H), 6.51 (s, 1H), 6.18 (s, 1H), 4.59 (d, J = 5.8 Hz, 2H), 4.50 (d, J =10.1 Hz, 2H), 4.34 (d, J = 9.5 Hz, 2H), 3.38 (s, 3H), 2.27 (d, J = 19.3Hz, 6H) 206 1 ¹H NMR (400 MHz, DMSO-d6) δ 11.83 (s, 1H), 9.19 526.3 (s,1H), 8.58 (d, J = 2.5 Hz, 1H), 8.41 (d, J = 2.2 Hz, 1H), 8.32 (d, J =7.6 Hz, 1H), 7.94 (dd, J = 8.5, 2.3 Hz, 1H), 7.87 (d, J = 8.4 Hz, 1H),7.79 (d, J = 1.7 Hz, 1H), 6.55 (dd, J = 2.6, 1.7 Hz, 1H), 5.15-5.06 (m,1H), 3.98 (d, J = 13.7 Hz, 2H), 3.43 (td, J = 9.5, 4.7 Hz, 2H), 2.87 (s,2H), 2.19 (s, 3H), 2.14-2.07 (m, 5H), 1.55-1.43 (m, 5H). 207 1 ¹H NMR(400 MHz, DMSO) δ 9.66 (s, 1H), 8.72 (t, J = 527 6.1 Hz, 1H), 8.60 (d, J= 2.5 Hz, 1H), 8.36 (s, 1H), 7.88 (d, J = 1.9 Hz, 2H), 7.81 (d, J = 1.0Hz, 1H), 6.57 (dd, J = 2.5, 1.7 Hz, 1H), 6.46 (s, 1H), 6.19 (s, 1H),4.36 (d, J = 6.1 Hz, 2H), 3.94 (d, J = 13.0 Hz, 2H), 3.25 (t, J = 11.2Hz, 2H), 3.17 (s, 3H), 2.26 (s, 3H), 2.20 (s, 3H), 2.02-1.93 (m, 2H),1.87 (d, J = 13.8 Hz, 2H). 208 1 ¹H NMR (400 MHz, DMSO) δ 9.50 (s, 1H),8.86 (t, J = 528 6.1 Hz, 1H), 8.68 (dd, J = 4.5, 0.6 Hz, 1H), 8.38 (s,1H), 7.95-7.86 (m, 3H), 6.86 (s, 1H), 6.62 (s, 1H), 6.15 (s, 1H), 5.92(s, 1H), 4.39 (d, J = 6.1 Hz, 2H), 4.21 (d, J = 9.1 Hz, 2H), 4.05 (d, J= 9.1 Hz, 2H), 3.32 (s, 3H), 2.22 (s, 3H). 209 1 ¹H NMR (400 MHz, DMSO)δ 11.59 (s, 1H), 11.00 (s, 529 1H), 8.48 (s, 1H), 8.36 (d, J = 1.7 Hz,1H), 7.84 (dd, J = 8.5, 2.2 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 6.34 (s,1H), 6.14 (s, 1H), 6.10 (s, 1H), 4.38 (d, J = 5.6 Hz, 2H), 4.16 (d, J =23.4 Hz, 2H), 3.22 (d, J = 10.1 Hz, 2H), 2.55 (s, 3H), 2.31 (s, 3H),2.25 (s, 3H), 2.20 (d, J = 7.9 Hz, 5H), 1.55 (q, J = 7.3 Hz, 2H), 1.46(t, J = 10.3 Hz, 2H), 0.70 (t, J = 7.4 Hz, 3H). 210 2 ¹H NMR (400 MHz,DMSO) δ 9.35 (s, 1H), 9.05 (d, J = 530 7.3 Hz, 1H), 8.69 (dd, J = 4.6,0.7 Hz, 1H), 8.45 (d, J = 2.2 Hz, 1H), 8.00 (dd, J = 8.6, 2.3 Hz, 1H),7.97-7.85 (m, 2H), 6.86 (s, 1H), 6.71 (d, J = 5.0 Hz, 1H), 6.12 (s, 1H),5.91 (s, 1H), 5.17-5.09 (m, 1H), 4.44-4.30 (m, 2H), 4.25-4.15 (m, 2H),2.19 (s, 3H), 1.50 (d, J = 7.1 Hz, 3H). 211 4 ¹H-NMR (400 MHz, DMSO-d6)δ ppm 11.63 (br. s., 530 1H), 8.68-8.66 (m, 1H), 8.53 (br. s., 1H), 8.31(d, 1H, J = 2.0 Hz), 7.82-7.73 (m, 2H), 6.34 (br. s., 1H), 6.09 (s, 1H),6.00 (s, 1H), 5.97 (s, 1H), 4.31 (d, 2H, J = 6.4 Hz), 3.96 (d, 1H, J =2.0 Hz), 3.16 (s, 3H), 3.11-3.05 (t, 2H, J = 12 Hz), 2.55 (s, 3H), 2.19(s, 3H), 2.17 (s, 3H), 2.14 (s, 3H), 1.88-1.75 (m, 4H). 212 1 ¹H NMR(400 MHz, DMSO) δ 9.66 (s, 1H), 8.78 (d, J = 531 8.1 Hz, 1H), 8.70-8.66(m, 1H), 8.45 (d, J = 2.1 Hz, 1H), 8.00 (dd, J = 8.6, 2.3 Hz, 1H), 7.91(dd, J = 13.3, 6.4 Hz, 2H), 6.35 (s, 1H), 6.22 (s, 1H), 5.09 (dd, J =14.8, 7.5 Hz, 1H), 4.44 (d, J = 9.5 Hz, 1H), 4.35 (d, J = 9.5 Hz, 1H),4.19 (dd, J = 14.2, 9.6 Hz, 3H), 3.27 (s, 3H), 2.20 (d, J = 3.9 Hz, 6H),1.49 (d, J = 7.1 Hz, 3H). 213 1 ¹H NMR (400 MHz, DMSO-d6) δ 11.88 (s,1H), 9.26 531 (s, 1H), 8.59 (d, J = 2.6 Hz, 1H), 8.49-8.25 (m, 2H), 7.88(d, J = 1.5 Hz, 2H), 7.84-7.71 (m, 1H), 6.54 (t, J = 2.1 Hz, 1H), 6.13(s, 2H), 4.35 (d, J = 5.7 Hz, 2H), 4.20 (d, J = 13.4 Hz, 2H), 3.13 (m,7H), 2.19 (s, 3H), 2.08 (m, 5H), 1.73 (t, J = 7.2 Hz, 2H), 1.40 (s, 2H).214 1 ¹H-NMR (400 MHz, CDCl3) δ ppm 8.47 (d, 1H, J = 2.4 531 Hz), 8.27(d, 1H, J = 1.2 Hz), 7.88 (d, 1H, J = 8.8 Hz), 7.67-7.65 (m, 2H), 6.69(br. s., 1H), 6.40 (d, 1H, J = 2.0 Hz), 6.09-6.07 (m, 1H), 5.98 (s, 1H),5.93 (s, 1H), 5.28- 5.26 (m, 1H), 4.33-4.29 (m, 2H), 3.35-3.26 (m, 3H),3.17 (s, 3H), 2.97 (s, 3H), 2.23 (s, 3H), 2.18 (s, 3H), 2.02-1.96 (m,4H), 1.50 (d, 3H, J = 6.8 Hz). 215 1 ¹H-NMR (400 MHz, DMSO-d6) δ ppm11.85 (s, 1H), 531 9.22 (br. s., 1H), 8.68-8.65 (m, 1H), 8.31 (d, 1H, J= 1.6 Hz), 7.80 (dd, 1H, J = 2.0, 8.4 Hz), 7.74 (d, 1H, J = 8.4 Hz),6.25-6.09 (m, 3H), 4.37-4.31 (m, 4H), 3.16-3.11 (m, 5H), 2.55 (s, 3H),2.19 (s, 6H), 2.11 (s, 3H), 1.79 (br. s., 4H). 216 4 ¹H NMR (400 MHz,DMSO-d6) δ 11.61 (s, 1H), 8.66 534.3 (d, J = 4.4 Hz, 1H), 8.50 (d, J =8.3 Hz, 2H), 8.41 (d, J = 2.3 Hz, 1H), 7.97 (dd, J = 8.6, 2.3 Hz, 1H),7.90 (d, J = 4.2 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 6.31 (s, 1H), 5.96(s, 2H), 5.04 (p, J = 7.2 Hz, 1H), 3.92 (dd, J = 20.0, 13.5 Hz, 2H),3.14 (s, 3H), 3.12-3.02 (m, 2H), 2.15 (s, 3H), 2.09 (s, 3H), 1.80 (dd, J= 28.3, 8.5 Hz, 4H), 1.45 (d, J = 7.0 Hz, 3H). 217 4 ¹H NMR (400 MHz,DMSO-d6) δ 8.66 (d, J = 4.5 Hz, 534.2 1H), 8.53 (s, 1H), 8.41 (d, J =2.3 Hz, 1H), 7.99 (d, J = 7.7 Hz, 1H), 7.95 (dd, J = 8.6, 2.3 Hz, 1H),7.89 (d, J = 4.3 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.25 (s, 1H), 5.98(s, 1H), 5.91 (s, 1H), 5.10 (q, J = 7.2 Hz, 1H), 4.90 (t, J = 5.2 Hz,1H), 3.83 (d, J = 13.2 Hz, 2H), 3.44 (d, J = 5.3 Hz, 2H), 2.99 (q, J =10.1 Hz, 2H), 2.16 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H), 1.51-1.38 (m,4H). 218 1 ¹H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 9.20 535 (s, 1H),8.66 (d, J = 4.5 Hz, 1H), 8.49 (d, J = 8.2 Hz, 1H), 8.41 (d, J = 2.2 Hz,1H), 8.05-7.71 (m, 3H), 6.15 (m, 2H), 5.04 (p, J = 7.2 Hz, 1H), 4.31 (t,J = 14.0 Hz, 2H), 3.21-3.08 (m, 5H), 2.18 (s, 3H), 2.10 (s, 3H), 1.90-1.63 (m, 4H), 1.46 (d, J = 7.1 Hz, 3H). 219 1 ¹H NMR (400 MHz, DMSO-d6)δ 11.82 (s, 1H), 9.15 535.2 (s, 1H), 8.66 (d, J = 4.5 Hz, 1H), 8.42 (d,J = 2.2 Hz, 1H), 8.03-7.91 (m, 2H), 7.91-7.80 (m, 2H), 6.11 (s, 2H),5.09 (t, J = 7.2 Hz, 1H), 4.89 (t, J = 5.3 Hz, 1H), 4.25-4.11 (m, 2H),3.44 (d, J = 5.3 Hz, 2H), 3.11 (q, J = 10.7 Hz, 2H), 2.18 (s, 3H), 2.08(s, 3H), 2.01 (d, J = 12.5 Hz, 2H), 1.50-1.32 (m, 5H). 220 2 ¹H NMR (301MHz, DMSO) δ 10.98 (s, 1H), 8.67 (d, J = 535 4.5 Hz, 1H), 8.62 (d, J =8.5 Hz, 1H), 8.42 (d, J = 2.1 Hz, 1H), 7.98 (dd, J = 8.6, 2.2 Hz, 1H),7.92 (d, J = 4.2 Hz, 1H), 7.87 (d, J = 8.6 Hz, 1H), 6.69 (s, 1H), 5.94(s, 1H), 5.04 (s, 1H), 4.53 (s, 1H), 3.99 (s, 1H), 3.37 (d, J = 38.2 Hz,2H), 3.18 (s, 4H), 2.36 (s, 4H), 2.25 (s, 4H), 1.98-1.79 (m, 5H), 1.47(d, J = 7.0 Hz, 4H). 221 3 ¹H NMR (400 MHz, DMSO-d6) δ 11.83 (s, 1H),9.18 536.2 (s, 1H), 9.11 (d, J = 1.4 Hz, 1H), 8.74-8.68 (m, 1H), 8.50(d, J = 1.4 Hz, 1H), 8.43 (d, J = 7.8 Hz, 1H), 8.05- 7.97 (m, 1H), 6.20(b.s., 1H), 6.10 (b.s., 1H), 5.12 (p, J = 7.1 Hz, 1H), 4.35-4.29 (m,2H), 3.19 (s, 3H), 3.17- 3.10 (m, 2H), 2.18 (s, 3H), 2.10 (s, 3H),1.87-1.67 (m, 4H), 1.48 (d, J = 7.0 Hz, 3H). 222 1 ¹H NMR (400 MHz,DMSO) δ 9.74 (s, 1H), 8.58 (dd, J = 541 10.0, 5.3 Hz, 2H), 8.43 (d, J =1.6 Hz, 1H), 7.98 (dd, J = 8.5, 2.0 Hz, 1H), 7.89 (d, J = 8.5 Hz, 1H),7.81 (s, 1H), 6.56 (s, 1H), 6.47 (s, 1H), 6.21 (s, 1H), 5.11-5.02 (m,1H), 3.91 (t, J = 13.7 Hz, 2H), 3.26 (t, J = 10.3 Hz, 2H), 3.16 (s, 3H),2.26 (s, 3H), 2.21 (s, 3H), 2.02-1.85 (m, 4H), 1.48 (d, J = 7.0 Hz, 3H).223 1 ¹H NMR (400 MHz, DMSO) δ 9.56 (s, 1H), 8.76 (d, J = 542 8.2 Hz,1H), 8.68 (d, J = 4.5 Hz, 1H), 8.44 (d, J = 2.1 Hz, 1H) 8 00 (dd, J =8.6, 2.2 Hz, 1H) 7.91 (dd, J = 12.8, 6.4 Hz, 2H), 6.93 (d, J = 55.8 Hz,1H), 6.60 (s, 1H), 6.14 (s, 1H), 5.92 (s, 1H), 5.14-5.06 (m, 1H), 4.24(s, 1H), 4.16 (s, 1H), 4.03 (d, J = 3.9 Hz, 2H), 3.29 (s, 3H), 2.22 (s,3H), 1.49 (d, J = 7.1 Hz, 3H). 224 1 ¹H NMR (400 MHz, DMSO-d6) δ 11.83(s, 1H), 9.19 544.2 (s, 1H), 8.66 (d, J = 4.5 Hz, 1H), 8.41 (d, J = 2.2Hz, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.95 (dd, J = 8.5, 2.3 Hz, 1H),7.92-7.82 (m, 2H), 5.14-5.06 (m, 1H), 4.04- 3.92 (m, 2H), 3.49-3.37 (m,2H), 2.87 (s, 2H), 2.19 (s, 3H), 2.10 (s, 5H), 1.54-1.43 (m, 5H). 225 4¹H-NMR (400 MHz, DMSO-d6) δ ppm 11.70 (s, 1H), 544 8.68 (d, 1H, J = 4.0Hz), 8.54 (br. s., 1H), 8.39 (d, 1H, J = 2.0 Hz), 8.10 (d, 1H, J = 8.0Hz), 7.95-7.87 (m, 3H), 6.25 (br. s., 1H), 5.97-5.92 (m, 2H), 5.09-5.06(m, 1H), 3.73-3.69 (m, 2H), 3.16-3.08 (m, 2H), 2.09 (s, 3H), 2.07-2.04(m, 2H), 1.84-1.80 (m, 1H), 1.48 (d, 3H, J = 7.2 Hz), 1.42-1.36 (m, 2H),1.15 (s, 3H), 0.91-0.89 (m, 2H), 0.64-0.61 (m, 2H). 226 1 ¹H NMR (400MHz, DMSO) δ 9.70 (s, 1H), 8.76-8.66 545 (m, 2H), 8.36 (s, 1H),7.97-7.83 (m, 3H), 6.48 (s, 1H), 6.20 (s, 1H), 4.35 (d, J = 6.0 Hz, 2H),3.94 (d, J = 13.0 Hz, 2H), 3.25 (t, J = 11.3 Hz, 2H), 3.17 (s, 3H), 2.26(s, 3H), 2.20 (s, 3H), 1.97 (t, J = 10.7 Hz, 2H), 1.87 (d, J = 13.5 Hz,2H). 227 1 ¹H NMR (400 MHz, DMSO) δ 9.74 (s, 1H), 8.67 (d, J = 559 4.4Hz, 1H), 8.57 (d, J = 8.2 Hz, 1H), 8.43 (d, J = 1.9 Hz, 1H), 7.99 (dd, J= 8.5, 2.1 Hz, 1H), 7.90 (dd, J = 15.5, 6.3 Hz, 2H), 6.47 (s, 1H), 6.21(s, 1H), 5.12-5.01 (m, 1H), 3.91 (t, J = 13.6 Hz, 2H), 3.26 (t, J = 10.0Hz, 2H), 3.16 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.02-1.84 (m, 4H),1.47 (d, J = 7.0 Hz, 3H). 228 1 ¹H NMR (400 MHz, DMSO-d6) δ 11.85 (s,1H), 9.22 571 (s, 1H), 8.64-8.45 (m, 2H), 8.34 (d, J = 1.8 Hz, 1H),7.96-7.82 (m, 2H), 7.79 (d, J = 1.6 Hz, 1H), 6.55 (t, J = 2.2 Hz, 1H),6.16 (m, 2H), 4.34 (d. J = 5.9 Hz, 2H), 4.31-4.18 (m, 2H), 4.00 (q, J =9.1 Hz, 2H), 3.61 (td, J = 6.6, 4.0 Hz, 1H), 3.21-2.99 (m, 1H), 2.19 (s,3H), 2.11 (s, 3H), 1.89 (s, 4H). 229 1 ¹H NMR (300 MHz, DMSO) δ 11.14(s, 1H), 9.02 (s, 575 1H), 8.46 (s, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.93(d, J = 8.4 Hz, 1H), 7.65 (s, 1H), 7.22 (s, 1H), 7.02 (s, 1H), 6.50 (s,1H), 6.18 (s, 1H), 4.47 (s, 2H), 4.45-4.40 (m, 2H), 4.34 (d, J = 10.5Hz, 2H), 3.35 (s, 3H), 2.30 (s, 3H), 2.24 (s, 3H).

Example 10. Assays

In order to assess the activity of chemical compounds against therelevant kinase of interest, the Caliper LifeSciences electrophoreticmobility shift technology platform was used. Fluorescently labeledsubstrate peptide was incubated in the presence of kinase and ATP sothat a reflective proportion of the peptide was phosphorylated. At theend of the reaction, the mix of phosphorylated (product) andnon-phosphorylated (substrate) peptides were passed through themicrofluidic system of the Caliper EZ Reader 2, under an appliedpotential difference. The presence of the phosphate group on the productpeptide provided a difference in mass and charge between those of thesubstrate peptide, resulting in a separation of the substrate andproduct pools in the sample. As the pools pass a LEDS within theinstrument, these pools were detected and resolved as separate peaks.The ratio between these peaks therefore reflects the activity of thechemical matter at that concentration in that well, under thoseconditions. The two specific assays used are described in detail below.

A. RET Wild Type Assay at KM

In each well of a 384-well plate, 7.5 nM-10 nM of wild type RET(ProQinase 1090-0000-1) was incubated in a total of 12.5 μL of buffer(100 mM HEPES pH 7.5, 0.015% BriJ 35, 10 mM MgCl₂, 1 mM DTT) with 1 μMCSKtide (FITC-AHA-KKKKD DIYFFFG-NH2) (SEQ ID NO: 5) and 25 μM ATP at 25°C. for 120 minutes in the presence or absence of a dosed concentrationseries of compound (1% DMSO final concentration). The reaction wasstopped by the addition of 70 μL of Stop buffer (100 mM HEPES pH 7.5,0.015% BriJ 35, 35 mM EDTA and 0.2% of Coating Reagent 3 (CaliperLifesciences)). The plate was then read on a Caliper EZReader 2(protocol settings: −1.7 psi, upstream voltage −500, downstream voltage−3000, post sample sip 35s). Data was normalized to 0% and 100%inhibition controls and the IC₅₀ calculated using a 4-parameter fit inthe CORE LIMS.

B. RET V804L Gatekeeper Mutant Assay at KM

In each well of a 384-well plate, 7.5 nM-10 nM of mutant RET (ProQinase1096-0000-1) was incubated in a total of 12.5 μL of buffer (100 mM HEPESpH 7.5, 0.015% BriJ 35, 10 mM MgCl2, 1 mM DTT) with 1 μM CSKtide(FITC-AHA-KKKKDDIYFFFG-NH2) (SEQ ID NO: 5) and 10 μM ATP at 25° C. for120 minutes in the presence or absence of a dosed concentration seriesof compound (1% DMSO final concentration). The reaction was stopped bythe addition of 70 μL of Stop buffer (100 mM HEPES pH 7.5, 0.015% BriJ35, 35 mM EDTA and 0.2% of Coating Reagent 3 (Caliper Lifesciences)).The plate was then read on a Caliper EZReader 2 (protocol settings: −1.7psi, upstream voltage −500, downstream voltage −3000, post sample sip35s). Data was normalized to 0% and 100% inhibition controls and theIC₅₀ calculated using a 4-parameter fit in the CORE LIMS.

In the Table 7 below, the following designations are used: <10.00 nM=A;10.01-100.0 nM=B; >100 nM=C; and ND=not determined.

TABLE 7 Compound Wild-Type V480L Mutant 100 C B 101 B B 102 C B 103 C C104 C B 105 B B 106 C B 107 A A 110 B A 111 A A 112 A A 113 B A 114 B B115 B B 116 C B 117 A A 118 A A 119 A A 120 A A 121 A A 122 A A 123 B B124 A A 125 C C 126 A A 127 A A 128 A A 129 A A 130 A A 131 A A 132 A A133 A A 134 A A 135 B B 136 A A 137 A A 138 B A 139 A A 140 A A 141 A A142 A A 143 A A 144 A A 145 A A 146 A A 147 B B 148 A A 149 A A 150 B B151 A A 152 A A 153 A A 154 C C 155 C B 156 A A 157 A A 158 A A 159 A A160 A A 161 A A 162 A A 163 B B 164 A A 165 A A 166 A A 167 A A 168 A A169 A A 170 A A 171 A A 172 A A 173 A A 174 A A 175 A A 176 B B 177 A A178 A A 179 A A 180 A A 181 A A 182 A A 183 A A 184 A A 185 A A 186 A A187 A A 188 A A 189 A A 190 A A 191 A A 192 A A 193 A A 194 A A 195 A A196 A A 197 A A 198 A A 199 A A 200 A A 201 A A 202 A A 203 B B 204 B A205 B A 206 A A 207 A A 208 A A 209 A A 210 A A 211 A A 212 A A 213 A A214 A A 215 A A 216 A A 217 A A 218 A A 219 A A 220 A A 221 A A 222 A A223 A A 224 A A 225 A A 226 A A 227 A A 228 A A 229 B B

Example 11. Compounds Disclosed Herein are Potent Inhibitors ofWild-Type and Mutant RET

In some embodiments, compounds according to structural Formula (I),(Ia), (II), and (111) are potent and selective inhibitors of oncogenicRET mutant and fusion proteins. Currently, there are no approvedinhibitors that selectively target these disease-driving RETalterations.

1. Additional In Vitro Assays

A compound described herein can be further tested in vitro forinhibition of other mutant RET kinases, including e.g., RET V804M andRET M918T kinases, as well as CCDC6-RET and KIF5B-RET fusion kinases.The IC₅₀ can be calculated.

2. Additional Cellular Assays

In cellular systems, the activity of a compound of structural Formula(I), (Ia), (II), or (III) can be measured by inhibition of RET mutant orRET fusion autophosphorylation, RET-dependent signaling, and byinhibition of RET dependent cell proliferation. More specifically, thecompound can be assayed for activity in cancer cell lines endogenouslyexpressing activated RET fusions or other mutants. Exemplary cells thatcan be used for these studies include patient xenografts and establishedcell lines. Exemplary patient xenografts include colorectal cancerpatient derived xenograft, Lung adenocarcinoma patient derivedxenograft, and NSCLC patient derived xenograft. Exemplary cell linesinclude Ba/F3-KIF5B-RET (a model for leukemia), LC2/ad cells (a modelfor lung cancer), MZ-CRC 1 (a model for thyroid cancer), and TT cells (amedullary thyroid cancer cell line). Exemplary RET mutations that can beused for these studies include fusions such as KIF5B-RET and CCDC6-RET;point mutations such as RET C634W, RET V804L, RET V804E, RET V804M, andRET M918T; and fusions containing point mutations such as KIF5B-RET(V804L) and KIF5B-RET (V804M). KIF5B-RET (V804L) refers to a mutant RETthat comprises a fusion with KIF5B and further comprises a V804Lmutation in RET (referring the amino acid numbering of wild-type RET).KIF5B-RET (V804M) refers to a mutant RET that comprises a fusion withKIF5B and further comprises a V804M mutation in RET.

2a. Autophosphorilation Assays

As an example of an autophosphorylation assay, in Ba/F3 modelsengineered to express KIF5B RET, the compound is tested for its abilityto inhibit RET fusion protein signaling as measured by inhibition of RETautophosphorylation. IC₅₀ can be calculated. Wild-type RET inhibitorscabozantinib and vandetanib can be used as controls (less potentcompounds than the test compound) in these cellular assays. In someembodiments, the compound of structural Formula (I), (Ia), (II), or(III) potently and selectively inhibit RET autophosphorylation.

The compound can also be tested for ability to inhibit RETautophosphorylation in LC2/ad cells, a non-engineered NSCLC cell linethat expressed a CCDC6-RET fusion (Suzuki et al, 2013 Cancer Sci. 104,896-903). For instance, immunoblot is performed for LC2/ad cellsexpressing the CCDC6-RET fusion protein and treated with the testcompound at different concentrations; Phosphorylated (phospho[Y1062])and total protein levels of RET are measured.

2b. Proliferation Assays

In some embodiments, the compound of structural Formula (I), (Ia), (II),or (III) inhibits proliferation.

In proliferation assays, the compound can be tested for ability toinhibit KIF5B-RET dependent Ba/F3 cell growth. The IC₅₀ can becalculated.

Inhibition of RET activity with the compound can also be tested forinhibition of proliferation of the CCDC6-RET expressing cell line.Similarly, the compound can also be tested for its ability to inhibitRET pathway signaling and RET dependent proliferation in the human MTCTT and MZ-CRC 1 cell lines, driven by RET C634W or RET M918T mutations,respectively. In some embodiments, in RET-driven cell lines, the testcompound inhibits RET activity and RET-driven proliferation morepotently than the multi-kinase inhibitors such as cabozantinib andvandetanib.

2c. Downstream Signalling Assays

In LC2/ad cells, a non-engineered NSCLC cell line that expressed aCCDC6-RET fusion (Suzuki et al, 2013), the compound can be tested forits ability to inhibit phosphorylation of the RET substrate Src homologydomain (Shc) (Hayashi et al, 2000 Oncogene. 19, 4469-4475), anddownstream signaling through extracellular signal regulated kinase(ERK)1/2, including downregulation of dual specificity phosphatase 6(DUSP6) and sprouty receptor tyrosine kinase signaling antagonist 4(SPRY4) (Lito et al, 2013 Nat. Med. 19, 1401-1409). For instance,immunoblot can be performed for LC2/ad cells expressing the CCDC6-RETfusion protein and treated with the test compound at differentconcentrations; phosphorylated and total levels of downstreambiomarkers, e.g., phospho(Y239/Y240)-Shc and phospho(Y202/T204)-ERK 1/2,are measured. In addition, to determine expression levels of downstreamtargets, LC2/ad cells can be treated with the compound, cabozantinib, orDMSO for 7 hours and RNA is harvested. Gene expression levels of DUSP6and SPRY4 can be measured by qRT-PCR. In some embodiments, the compoundinduces a dose-dependent decrease in expression of the ERK1/2 targetgenes DUSP6 and SPRY4 but not the control gene glycogen synthase kinase3 beta (GSK3B).

3. Animal Models

Antitumor efficacy of compounds of structural Formula (I), (Ia), (II),or (III) can be demonstrated in several RET-driven in vivo models. TheBa/F3-KIF5B-RET allograft model uses KIF5B RET fusion-dependent Ba/F3cells. The test compound can be administered orally with an appropriatedose. Tumor size can be measured e.g., twice weekly. In someembodiments, administration of the compound results in robust anddose-dependent growth inhibition of Ba/F3-KIF5B-RET allograft tumors,e.g., in complete TGI and Mouse body weight can be measured e.g.,twice-weekly during the administration. In some embodiments, thecompound is well tolerated with no significant changes in animal bodyweight observed.

Similar assays can be performed using other animal models, including aBa/F3-KIF5B-RET (V804L) allograft tumor model which comprises a KIF5BRET V804L fusion protein, an KIF5B-RET NSCLC allograft tumor model, anMTC cell line xenograft driven by a RET C634W mutation, and a CCDC6-RETfusion positive colorectal cancer allograft tumor model. The RET V804Lmutation has been observed in rare cases of MTC and is predicted to beinsensitive to cabozantinib and vandetanib in vitro and in vivi(Carlomagno et al, 2004 Biochem. Biophys. Res. Commun. 207, 1022-1028;Bentzien et al, 2013 Thyroid. 23, 1569-1577). In some embodiments, thecompound causes complete TGI and regressions in a cancer that is notresponsive to cabozantinib or vandetanib.

Biochemical markers can also be assayed in the treated mice. To assessdirect inhibition of KIF5B-RET (V804L) fusion kinase activity in Ba/F3KIF5B-RET (V804L) tumors, the compound can be administered orally at anappropriate dose to tumor bearing mice for several days and plasma andtumors can be collected from individual mice at appropriate time pointsafter the last dose. Test compound concentrations in plasma can bedetermined by liquid chromatograph/tandem mass spectrometry (LC/MS/MS).Inhibition of KIF5B-RET (V804L) signaling in the tumor tissue can beassessed by a phosphor RET enzyme linked immunosorbent assay (ELISA) andby immunoblotting, e.g., as described above. Quantitation of thephospho-RET signal by ELISA can measure the percent KIF5B-RET (V804L)inhibition in treated animals as compared to vehicle treated controls.Suppression of downstream RET pathway signaling can be demonstrated byinhibition of Shc phosphorylation. In some embodiments, a dose andtime-dependent correlation is observed between the concentration of thetest compound in mouse plasma and the level of phosphorylated KIF5B RET(V804L). In some embodiments, administration of the compound at anamount sufficient to reach at least 90% inhibition of RET in vivo leadsto therapeutic efficacy, e.g., can lead to 100% tumor growth inhibition.

Example 12. Selectivity of Compounds of Structural Formulas (I), (Ia),(II), and (III)

Efficacy Against Wild-Type and Mutant RET

In some embodiments, compounds according to structural Formula (I),(Ia), (II), and (III) are potent inhibitors of wild-type and mutant RET.For instance the IC₅₀ of a compound can be tested in a cell linecomprising wild-type RET and in a second cell line comprising mutantRET, e.g., a point mutation or fusion.

Selectivity for RET Over KDR

In some embodiments, compounds according to structural Formula (I),(Ia), (II), and (III) are selective for RET over another kinase, such asKDR (also called Vascular endothelial growth factor receptor 2). KDR isa tyrosine-protein kinase that acts as a cell-surface receptor forVEGFA, VEGFC and VEGFD. Inhibition of KDR/VEGFR2 has been associatedclinically with certain adverse effects, e.g., hypertension, arterialthrombosis, and hemorrhage, and therefore selectivity for RET over KDRis desirable.

To test selectivity, the test compound can be assayed for its ability toinhibit proliferation in parental Ba/F3 cells that do not express a RETmutation, e.g., does not express a KIF5B-RET fusion. A weak IC₅₀ in theparental cell line indicates that the test compound is selective forcell lines dependent on oncogenic RET.

The selectivity of a compound on RET versus other human kinases can becharacterized by profiling binding across a panel of over 450 humankinases and disease-relevant kinase mutants. In some embodiments, thecompound has a high degree of selectivity for RET and RET kinase mutantsover other kinases tested. To define the binding affinity for thekinases bound by the compound in kinome screening and additional kinasesof interest, the dissociation constant (Kd) can be determined.

To differentiate the compound from multi-kinase inhibitors withbiochemical activity against RET, the activity of the compound againstrecombinant kinase insert domain receptor (KDR) (also known as vascularendothelial growth factor receptor 2 [VEGFR2]) and fibroblast growthfactor receptor 1 (FGFR1) can be tested, as inhibition of these kinasesis associated with dose-limiting toxicities in humans. Inhibition ofKDR/VEGFR2 has been associated clinically with hypertension, arterialthrombosis, and hemorrhage, whereas inhibition of fibroblast growthfactor receptors (FGFRs) is associated with hyperphosphatemia and tissuecalcification (Abdel-Rahman and Fouad, 2014 Crit. Rev. Oncol. Hematol.92, 194-207; Touat et al, 2015 Clin. Cancer Res. 21, 2684-2694). In someembodiments, the compound is a more potent inhibitor of WT RET thanKDR/VEGFR2 and FGFR1, respectively. In contrast, in some embodiments amulti-kinase inhibitor exhibits approximately equal or increased potencyon KDR versus WT RET.

Example 13. Selective Compounds Prevent RET Resistance Mutants

The compounds herein can be tested for the propensity of a cancer todevelop one or more RET mutations associated with drug resistance. Forexample, RET-altered cancer cells (e.g., Ba/F3 KIF5B-RET cells) can betreated with a mutagen such as ENU, exposed to a compound herein or acontrol compound (e.g., for 2-3 weeks), and the cell number can bequantified. Cells with high proliferation can be subjected to DNAsequencing to detect RET mutations. In some embodiments, a compound ofstructural Formula (I), (Ia), (II), or (III) leads to no or fewer RETmutations than a control compound such as a multi-kinase inhibitor suchas cabozantinib.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated byreference in their entirety as if each individual publication or patentwas specifically and individually indicated to be incorporated byreference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the disclosure described herein. Such equivalents areintended to be encompassed by the following claims.

1. A compound of structural formula (I) or a pharmaceutically acceptablesalt thereof, wherein:

ring A is an aryl or heteroaryl ring; each of X¹ and X² is independentlyselected from N and C(R⁶); Z is

—CD(R⁵)—, or —CH(R⁵)—, wherein “1” represents a point of attachment toN(R⁸); and “2” represents a point of attachment to ring A; each R¹ andeach R⁷ is independently selected from C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₁-C₆ alkoxy, halo, C₁-C₆ heteroalkyl, cycloalkyl, aryl,heteroaryl, aryloxy, aralkyl, heterocyclyl, heterocyclylalkyl, nitro,cyano, —C(O)R^(c), —OC(O)R^(c), —C(O)OR^(d), —(C₁-C₆alkylene)-C(O)R^(c), —SR^(d), —S(O)₂R^(c), —S(O)₂—N(R^(d))(R^(d)),—(C₁-C₆ alkylene)-S(O)₂R^(c), —(C₁-C₆ alkylene)-S(O)₂—N(R^(d))(R^(d)),—N(R^(d))(R^(d)), —C(O)—N(R^(d))(R^(d)), —N(R^(d))—C(O)R^(c),—N(R^(d))—C(O)OR^(c), —(C₁-C₆ alkylene)-N(R^(d))—C(O)R^(c),—N(R^(d))S(O)₂R^(c), and —P(O)(R^(c))(R^(c)), wherein each of alkyl,alkenyl, alkynyl, alkoxy, heteroalkyl, cycloalkyl, aryl, heteroaryl,aryloxy, aralkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a), or two R¹ or two R⁷ are takentogether with the carbon atoms to which they are attached form acycloalkyl or heterocyclyl ring independently substituted with 0-5occurrences of R^(b); each of R², R³ if present, and R⁴ is independentlyselected from hydrogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, halo, hydroxyl,cyano, C₁-C₆ heteroalkyl, and —N(R^(d))(R^(d)); wherein each of alkyl,alkoxy, and heteroalkyl is independently substituted with 0-5occurrences of R^(a); each of R⁵ and R⁸ is independently selected fromhydrogen, deuterium, C₁-C₆ alkyl, and C₁-C₆ heteroalkyl, wherein eachalkyl and heteroalkyl is independently substituted with 0-5 occurrencesof R^(a); each R⁶ is independently selected from hydrogen, C₁-C₆ alkyl,C₁-C₆ alkoxy, halo, cyano, C₁-C₆ heteroalkyl, and —N(R^(d))(R^(d)),wherein each alkyl, alkoxy, and heteroalkyl is independently substitutedwith 0-5 occurrences of R^(a); each R^(a) and each R^(b) isindependently selected from C₁-C₆ alkyl, halo, hydroxyl, C₁-C₆heteroalkyl, C₁-C₆ alkoxy, cycloalkyl, heterocyclyl, and cyano, whereineach of alkyl, heteroalkyl, alkoxy, cycloalkyl, and heterocyclyl isindependently substituted with 0-5 occurrences of R′; each R′ isindependently selected from C₁-C₆ alkyl, C₁-C₆ heteroalkyl, halo,hydroxyl, cycloalkyl, and cyano, or two R′ together with the atom(s) towhich they are attached form a cycloalkyl or heterocyclyl ring; eachR^(c) is independently selected from hydrogen, hydroxyl, halo, thiol,C₁-C₆ alkyl, C₁-C₆ thioalkyl, C₁-C₆ alkoxy, C₁-C₆ heteroalkyl,cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, wherein each of alkyl, thioalkyl, alkoxy,heteroalkyl, cycloalkyl, cycloalkylalkyl, heteroarylalkyl, heterocyclyl,and heterocyclylalkyl is independently substituted with 0-5 occurrencesof R^(a), or two R^(c) together with the atom(s) to which they areattached form a cycloalkyl or heterocyclyl ring independentlysubstituted with 0-5 occurrences of R^(b); each R^(d) is independentlyselected from hydrogen, C₁-C₆ alkyl, C₁-C₆ heteroalkyl, cycloalkyl,cycloalkylalkyl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl,wherein each of alkyl, heteroalkyl, cycloalkyl, cycloalkylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl is independentlysubstituted with 0-5 occurrences of R^(a), or two R^(d) together withthe atom(s) to which they are attached form a cycloalkyl or heterocyclylring independently substituted with 0-5 occurrences of R^(b); m is 0, 1,or 2; and n is 0, 1, 2, or
 3. 2. The compound of claim 1 or apharmaceutically acceptable salt thereof having the structural formula(Ia):

wherein: R¹ is halo, C₁-C₄ alkyl, C₃-C₆ cycloalkyl, or C₁-C₆ alkoxy; andR¹ is substituted with 0-3 occurrences of R^(a).
 3. The compound ofclaim 1 or a pharmaceutically acceptable salt thereof, wherein: R² isselected from hydrogen, C₁-C₄ alkyl substituted with 0-5 occurrences ofR^(a), C₁-C₆ alkoxy substituted with 0-5 occurrences of R^(a), hydroxyl,and halo; R³ if present is hydrogen; or R⁴ is selected from hydrogen,hydroxyl, halo, cyano, C₁-C₄ alkyl, and O—C₁-C₄ alkyl, wherein eachalkyl portion of R⁴ is substituted with 0-3 occurrences of R^(a).
 4. Thecompound of claim 3 or a pharmaceutically acceptable salt thereof,wherein: R² is hydrogen, fluoro, —CH₃, —CH₂CH₃—CH₂OH, —CH₂CN, —OCH₂CF₃,—OCH₂CH₂, or OMe; or R⁴ is selected from hydrogen, fluoro, cyano,hydroxyl, —CH₃, —CH₂CN, —CH₂CH, —CH₂CH₂OCH₃, —OCH₃, —OCH₂CF₃, and—OCH₂CH₃.
 5. The compound of claim 1 or a pharmaceutically acceptablesalt thereof, wherein Z is selected from

—CH₂—, and —CH(C₁-C₄ alkyl)-, wherein the C₁-C₄ alkyl is substitutedwith 0-3 occurrences of R^(a).
 6. The compound of claim 1 or apharmaceutically acceptable salt thereof, wherein: each R⁶ isindependently selected from hydrogen, halo, cyano, and C₁-C₄ alkylsubstituted with 0-3 occurrences of R^(a); or R⁸ is hydrogen and —CH₃.7. The compound of claim 1 or a pharmaceutically acceptable saltthereof, wherein ring A is phenyl or a 6-membered monocyclic heteroarylcomprising at least one nitrogen ring atom.
 8. The compound of claim 1or a pharmaceutically acceptable salt thereof, wherein ring A isselected from


9. The compound of claim 1 or a pharmaceutically acceptable saltthereof, wherein: ring A is selected from

n is 1; R⁷ is selected from 1H-pyrazol-1-yl, azetidin-1-yl, andpyrrolidin-1-yl; and R⁷ is substituted with 0-3 occurrences of R^(a).10. The compound of claim 9 or a pharmaceutically acceptable saltthereof, wherein R⁷ is selected from 3-fluoroazetidin-1-yl,3,3-difluoropyrrolidin-1-yl, 3-fluoropyrrolidin-1-yl,3-difluoromethyl-1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,4-chloro-1H-pyrazol-1-yl, 3-difluoromethyl-1H-pyrazol-1-yl,4-difluoromethyl-1H-pyrazol-1-yl, 4-cyclopropyl-1H-pyrazol-1-yl,4-fluoro-1H-pyrazol-1-yl, 3,5-bis(difluoromethyl)-1H-pyrazolyl,3-methyl-1H-pyrazol-1-yl, 4-methyl-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyrazol-1-yl.
 11. A compound havingthe structural formula (II):

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and C(R¹³); each Y¹ and Y² is independently selected from N andCH, wherein no more than one of Y¹ and Y² is N; Q is selected from N,CH, and CH₂; R¹¹ is C₁-C₄ alkyl; R¹² is selected from hydrogen and C₁-C₄alkyl; R¹³ if present is selected from hydrogen, cyano, and halo; R¹⁴ isselected from hydrogen, halo, cyano, hydroxyl, C₁-C₄ alkyl, and C₁-C₄alkoxy; R¹⁵ is selected from hydrogen and C₁-C₄ alkyl; R¹⁶ is selectedfrom hydrogen, and C₁-C₄ alkyl optionally substituted with 1 or moreindependently selected halo; R¹⁷ is selected from hydrogen and C₁-C₄alkyl; each of R^(18a) if present, R^(19a) if present, R^(18b), andR^(19b) is independently selected from hydrogen, halo, C₁-C₄ alkyloptionally substituted with one or more halo, and C₃-C₆ cycloalkyl; p is0 or 1; and each

represents a single or a double bond.
 12. The compound of claim 11 or apharmaceutically acceptable salt thereof, wherein: R¹¹ is —CH₃; R¹² isselected from hydrogen and —CH₃; R¹³ if present is selected fromhydrogen, cyano and fluoro; R¹⁴ is selected from hydrogen, fluoro,cyano, hydroxyl, —CH₃, —CH₂CH₃, —OCH₃, and —OCH₂CH₃; R¹⁵ is selectedfrom hydrogen and —CH₃; R¹⁶ is selected from hydrogen, —CH₃ and —CHF₂;R¹⁷ is selected from hydrogen and —CH₃; each of R^(18a) and R^(19a) ifpresent is independently selected from hydrogen and fluoro, wherein atleast one of R^(18a) or R^(19a) is hydrogen; each of R^(18b) and R^(19b)is independently selected from hydrogen, fluoro, chloro, —CH₃, —CHF₂,and cyclopropyl, wherein at least one of R^(18b) or R^(19b) is hydrogen;and each

is the same.
 13. A compound having structural formula (III):

or a pharmaceutically acceptable salt thereof, wherein: X¹ is selectedfrom N and CH; Z′ is selected from

or —CH(R²⁸)—, wherein “1” represents a point of attachment to N(R²⁶);and “2” represents a point of attachment to ring B; ring B is selectedfrom phenyl, pyridinyl, 1H-pyrazolyl, and pyrazinyl; R²¹ is selectedfrom C₃-C₆ cycloalkyl and C₁-C₄ alkyl; R²² is selected from hydrogen andC₁-C₄ alkyl; R²³ is selected from hydrogen and cyano; R²⁴ is selectedfrom hydrogen, hydroxy, and halo; R²⁵ is selected from hydrogen, halo,hydroxy, C₁-C₄ alkoxy, —C₁-C₄ alkyl, —C₁-C₄ alkyl-O—C₁-C₄ alkyl, whereineach C₁-C₄ alkyl is optionally substituted with 1 or more substituentsindependently selected from halo and cyano; R²⁶ is selected fromhydrogen and C₁-C₄ alkyl; R²⁷, if present, is independently selectedfrom 1H-pyrazolyl, pyridinyl, and C₁-C₄ alkoxy, wherein the1H-pyrazol-1-yl is optionally substituted with up to 2 substituentsindependently selected from C₁-C₄ alkyl and halo; R²⁸ is selected fromhydrogen and C₁-C₄ alkyl; and o is 0 or
 1. 14. The compound of claim 13or a pharmaceutically acceptable salt thereof, wherein: Z is selectedfrom

—CH₂, or —CH(CH₃)—; the portion of the molecule represented by

is selected from

R²¹ is selected from —CH₃ and cyclopropyl; R²² is selected from hydrogenand —CH₃; R²³ is selected from hydrogen and cyano; R²⁴ is selected fromhydrogen, hydroxy and fluoro; R²⁵ is selected from hydrogen, fluoro,hydroxy, —OCH₃, —OCH₂CF₃, —CH₂CH₂OCH₃, —CH₃, —CH₂CH₃, and —CH₂CN; R²⁶ isselected from hydrogen and —CH₃; and R³⁷ is selected from hydrogen,—OCH₃, —OCH₂CH₃, 1H-pyrazol-1-yl, 4-fluoro-1H-pyrazol-1-yl,3,5-dimethyl-1H-pyrazol-1-yl, and pyridin-2-yl.
 15. A pharmaceuticalcomposition comprising a compound of claim 1 or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier. 16.A method for inhibiting RET activity in a cell or in a patient,comprising the step of contacting said cell or administering to saidpatient a compound of claim 1 or a pharmaceutically acceptable saltthereof.
 17. A method for treating a patient suffering from a conditionmediated by aberrant RET activity, comprising administering to saidpatient a therapeutically effective amount of a compound of claim 1 or apharmaceutically acceptable salt thereof.
 18. A method for treating apatient who has developed resistance to a treatment for a conditionmediated by aberrant RET activity, comprising administering to saidpatient a therapeutically effective amount of a compound of claim 1 or apharmaceutically acceptable salt thereof.
 19. A method of treatingcancer in a patient in need thereof, said method comprising: a.determining if, having determined if, or receiving information that saidpatient has a RET-altered cell, cancer, gene, or gene product; b.identifying the said patient as responsive to a compound of claim 1 or apharmaceutically acceptable salt thereof when said patient has theRET-altered cell, cancer, gene, or gene product; and c. administering aneffective amount of the compound to said patient.
 20. A method oftreating cancer in a patient, said method comprising administering aneffective amount of a compound of claim 1 or a pharmaceuticallyacceptable salt thereof to a patient having a RET-altered cell, cancer,gene, or gene product that is responsive to the compound.